Reference Intervals for CD4 and hemoglobin among apparently healthy pregnant women in Addis Ababa, Ethiopia

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This is not appropriate because CD4 cell count tests are affected by ethnic and environmental factors Therefore, this study aimed to develop RI for CD4 cell count and hemoglobin levels for apparently healthy pregnant women in Addis Ababa, Ethiopia. Method From January to June 2019, 162 pregnant women who appeared healthy were enrolled in this community-based cross-sectional study. Social and medical data were gathered through a structured questionnaire following the acquisition of informed consent. BD FACS Presto, a near-patient CD4 counter, was used to obtain the CD4 count and hemoglobin (Hgb) levels from whole blood using a nonparametric test that estimated 2.5th (lower limit) and 97.5th (upper limit) percentiles with 95% CIs. Data were entered and analyzed using SPSS 20 and P < 0.05 was considered statistically significant. Results : After excluding six pregnant women who did not pass the screening tests, 156 apparently healthy pregnant women participants who were 18–49 years old were included in the final analysis. The majority of them were in the 2nd trimester, from the total participants (51.9%) were housewives, and (37.2. %) attained at least primary-level education. The medians of CD4%-and CD4# counts with inter-quartile ranges [IQR] were 757.5 [611.3-925.5] cells/µLand 43.62% [39.9–47.3], respectively while the median and IQR. Hemoglobin values were 14.3gm/dL [13.40–15.10]. The absolute CD4 cell count reference intervals were 416.9-1218.4 cells/µL, %CD4 + 32.1–57.3%, there is no significant difference between the trimester and age difference was observed. Conclusion : Comparing this study's results to those of non-African and African nations, it revealed a decline in both parentage count and absolute CD4 + T cell counts. Therefore, it is essential to establish local reference values for various populations. CD4 count %CD4 Hemoglobin Reference Interval Ethiopia Figures Figure 1 Introduction CD4 cells are a subset of T-lymphocytes that act as helper or inducer T-cells. Because they don’t kill the pathogen directly instead they induce a signal to other T cells and macrophages to initiate an immune response. The Cell is to mediate adapting immunity for the different pathogens, immunological memory and also participate in autoimmunity, asthma, and allergic responses when during individual become susceptible to a wide range of infectious disorders. ( 1 , 2 ) During pregnancy, some physiological changes occur that influence immunological activities that change the CD4 count. It provides a figure of the immune system in health; a higher CD4 count typically indicates the wellness of the immune system condition. ( 3 , 4 , 5 ) CD4 count can be affected by different factors starting from sample collection, up to analysis those are pathological, and physiological factors such as the technique and time of blood collection, transport, and storage of the specimens, posture, exercise, methodology, and instrument used to obtain the result ( 6 ) many laboratories lack RI for pregnant women, even though the physiological and biochemical changes in pregnancy influence many of our laboratory tests. Without adequate RI, there is an increased risk of missing significant changes due to pathological conditions and incorrectly interpreting normal changes as pathological events( 7 ) Certain laboratories have identified this condition and provide RI to pregnant women, even though pregnancy itself causes the same changes in routine laboratory tests. The majority of labs are actually unaware of the typical RI for expectant mothers. The results change depending on the trimester of pregnancy; for instance, the first and last trimesters have the highest hemoglobin concentrations, while the second trimester has the lowest. ( 8 ) In comparison to populations in some other African nations and the West, adult Ethiopian populations have reportedly been found to have higher hemoglobin levels. Conversely, during pregnancy, which is marked by numerous changes in practically all organ systems to accommodate the growing and developing fetal placenta, they have a lower CD4 cell count and CD4+/CD8 + T-cell ratio ( 9 ). Units, leading to a reduction in the majority of laboratory test parameters. ( 10 , 11 ) According to CLSI recommendation, each laboratory shall establish its reference range because the RI is affected by several factors including ethnicity, nutrition, altitude, geographic location, and pregnancy ( 12 ) Therefore, this study was designed to develop CD4 cell count and hemoglobin RI in apparently healthy pregnant women in Addis Ababa which was important as a guide for clinical support and treatment monitoring of patients in disease conditions and clinical trials. The findings will also be used as a reference for future studies. Materials and methods Study design, period, and setting For voluntary, seemingly healthy pregnant women in Addis Ababa, Ethiopia, a community-based cross-sectional study was conducted from January to June 2019. Addis Ababa lies between 2200 and 2500 meters over ocean level. It was the establishment of distinctive population bunches moreover known as the biggest capital city of the nation. And covers 527 square kilometers of zone in Ethiopia, and it has a 3,384,569 population size concurring to the 2007 census ( 13 ) during the study period, the city was categorized into 10 sub-cities. Based on the population estimate, 4 sub-cities (Yeka, Akaki Kality, Arada, and Kirkos) were chosen. It appears that the study included healthy pregnant women in Addis Ababa who were between the ages of 18 and 49 and who satisfied the eligibility requirements. Anemia, chronic renal failure, hypertension, coronary heart disease, diabetes, thyroid disease, liver disease, or any cancer is examples of chronic illnesses. People who use pharmacologically active substances aside from iron and who have received a blood transfusion within the last year are also listed. Additionally, factors like intestinal parasites, malaria, high CRP levels, smoking, drinking, and supplementing with folic acid were disregarded. The sampling strategy used in this study was a probability ratio. The sample size was established following the CLSI recommendation that a good way to develop RI is to collect samples from a sufficient number of different individuals to obtain at least 120 samples for analysis. PPS, where size is determined by the number of households in the city's Woredas (previously Kebeles, the smallest administrative units). Due to the size of Addis Abebe, four suburbs Arada, Kirkos, Akaki, and Yeka were chosen for the PPS; as a result, all of the chosen suburban woredas were included. Using Woreda as the sampling frame, systematic random sampling was used to select study participants. Informed consent was obtained before a blood sample was taken for laboratory analysis, and structured questionnaires were utilized to gather socio-demographic information. Specimen collection and laboratory analysis Using a vacutainer tube filled with ethylene diamine tetra acetic acid (EDTA) tubes, the sample was collected. Samples were taken prior to 11:00 a.m. to prevent daily variations and are identified with distinct codes. At the National HIV Reference Laboratory of the Public Health Institute of Ethiopia, a point-of-care FACS Presto (BD, San Jose, CA, USA) was used to analyze the CD4 count and hemoglobin in both absolute and percentage terms within eight hours of sample collection. Since 2016, Ethiopia has been using BD FACS PrestoTM CD4 point-of-care technology, which has a contract with FACS Calibur that is acceptable for CD4 cell counts and CD4 ( 14 ). The laboratory analyses were conducted strictly in accordance with standard operating procedures in order to guarantee the quality of the results. Furthermore, the external quality system is a part of the National Reference Laboratory. Statistical Analysis SPSS 20 was used to enter and analyze the data. With SPSS 20, the data distribution was evaluated and the data were input and examined. After the distribution of the data was evaluated, the data were compiled, summarized, and shown as absolute numbers with percentages for categorical variables and mean ± SD and medians with IQR values for continuous variables. The nonparametric Spearman correlation test was used to measure correlation, and the nonparametric Manutiny U and Kruskal-Wallis tests were used to compare groups. A non-parametric test was used to calculate RIs, estimating percentages of 2.5, the lower limit, and 97.5, the upper limit, with a 95% confidence interval. The RI was determined to be the 95 percent interval. It was determined that 0.05 was statistically significant. Ethical Considerations Regarding ethical issues, The Helsinki Declaration was followed in the conduct of this study. The Departmental Research and Ethics Review Committee of Addis Ababa University's Department of Medical Laboratory Science examined and approved the study protocol. At the time of enrollment, consent was obtained from voluntary participants, and all participants gave written informed consent. Participants received their CD4 + and hemoglobin test results back upon request, utilizing the BD FACSPresto. Results Socio-demographic characteristics 162 apparently healthy pregnant women aged 18–49 years were included in the determination of CD4 cells and hemoglobin RI. Of these, 6 participants were excluded due to chronic diseases, parasitic infections, HIV, and high CRP. A total of 156 participants participated in the final analysis. The median age of the study participants was 27 (IQR = 24–29) years. More than half (58.33%) of the participants were 25–34 years old. Most of the participants (88.4%) were married, 37.2% had primary school education and 51.9% were housewives (Table 1 ). Table 1 Socio-demographic characteristics of study participants in Addis Ababa, Ethiopia from January to June 2019 Variables Frequency Percentage (%) Sex Female 156 100 Age(years) 18–24 57 36.54 25–34 91 58.33 35–49 8 5.13 Marital status Single 16 10.3 Married 138 88.4 Divorced 2 1.3 Educational status Illiterate 12 7.7 Read and write 5 3.2 Primary 58 37.2 Secondary 57 36.5 College and above 24 15.4 Occupation Housewife 81 51.9 Government employee 14 9.0 Private employee 61 39.1 CD4 + T-cells and Hemoglobin by study participant’s characteristics The median value of CD4 count and Hbg concentration by different groups of study participant’s characteristics is presented in Table 2 . Overall status there was no statistically significant difference in CD4 count and hemoglobin concentration with participant’s characteristics except hemoglobin concentration in the gestation stage (trimester). The median value of absolute CD4 value was 779 (640–937) cells/µl 725 (594-927.5)cells/µl and 779(619.5–916) cells/µl in the first, second, and third trimesters respectively. The hemoglobin concentration of participants in the first trimester (median: 14.9, IQR: 14.3–15.4) is significantly higher than that of participants in the second (median: 14.1, IQR: 13.3–14.9) and third ((median: 14.1, IQR: 13.2–15.0) trimesters (p < 0.05). The hemoglobin concentration of pregnant women is inversely proportional to gestational age. Table 2 Median (IQR) value of CD4 count and hemoglobin concentration by different characteristics of study participants Addis Ababa, Ethiopia from January to June 2019 Characteristics (N = 156) AbsoluteCD4 count (cells/µL) CD4% Hemoglobin (g/dL) Median (IQR) Median(IQR) Median(IQR) Age(Years) 18–24 682 (594–875) 42.6 (39.4–46.0) 14.3(13.2–15.1) 25–34 794 (621.5–961) 44.2(40.1–48.8 14.3(13.5–15.2 35–49 730 (590–925) 41.8(38.1–44.8 15.2(13.4–15.4) Gestation stages (Trimester) 1st (n = 33) 779 (640–937) 43.2(39.0-47.7) *14.9(14.3–15.4) 2nd (n = 49) 725(594-927.5) 43.4(40.2–48.8) 14.1(13.3–14.9) 3rd (n = 74) 779(619.5–916) 43.8(40.0-46.8) 14.1(13.2–15.0) Education status Illiterate 726 (606–859) 43.9(41.1–46.4) 16.7(13.6–15.4) Read and write 931(750.5-1101.5) 46(37.0-47.4) 15.3(14.6–16.4) Primary 791.5(619.0-934.0) 44(40.1–47.1) 14.4(13.7–15.2) Secondary 723(565.5–899.0) 42.5(39.1–47.0) 12.1(13.2–14.9) College and above 804.5(719.3-922.8) 45.3(41.3–50.7) 14.5(13.2–15.3) Occupational status Housewife 750.5(608–930) 43.5(40.2–46.4) 14.3(13.5–15.2) Government employee 804.5(701–1029) 44.2(40.2–50.7) 14.7(13.4–15.4) Private employee 761(601–911) 43.8(39.3–48.8) 14.3(13.3–15.1) BMI Underweight 731.5(629.3-867.5) 37.2(36.0-52.3) 13.1(12.9–14.0) Healthy Weight 736.5(596.3–897) 43.5(40.1–47.7) 14.2(13.4–15.1) Overweight 775(627.8–983) 43.4(39.8–46.4) 14.5(13.6–15.2) Obesity 918(705.5–1118) 45.9(41.8–53.3) 15.0(13.9–15.6) Folate supplementation Supplemented 768.5(594.8-929.3) 43.1(39.7–46.3) 14.0(13.2–14.8) Not Supplemented 746.5(615.0-926.3) 44.2(40.0-48.2) 14.4(13.5–15.2) Iron supplementation Supplemented 768.5(613.0-925.5) 43.7(40.2–47.4) 14.3(13.4–15.2) Not Supplemented 719.5(565.8-927.5) 43.0(37.8–46.9) 14.8(13.0-15.3) *p = 0.018 Correlation of CD4 count and hemoglobin concentration with gestational age We assessed the correlation of absoluteCD4 count, CD4% and hemoglobin concentration with gestational age (Fig. 1). No significant correlation with gestational age for both CD4 absolute count (r=-0.042; p = 0.605) and CD4% (r=-0.006; p = 0.944) observed, while Hemoglobin concentration had weak negative correlation with gestational age (r=-0.160; p = 0.048). Figure1 - Correlation of CD4 absolute count (cells/µL), CD4% and hemoglobin concentration (g/dL) with gestational age (week) in pregnant women Correlation of A) absoluteCD4 count Vs gestational age, B) CD4% Vs gestational age and C hemoglobin Vs gestational age. Non-parametric spearman correlation test used to assess correlation. Absolute and Percentage CD4cells and Hemoglobin Reference Intervals AbsoluteCD4 count ranged between 394–1699 cells/µL while CD4% ranged 22–63% and Hgb 11–18 g/dL. According to Table 3 , the ranges for CD4% and absolute CD4 + T-cell counts were 32.1–57.3% and 416.9–1218.4 cells/µL, respectively. Table 3 Mean, Median, Range and 95% reference interval of CD4 and hemoglobin for healthy pregnant women in Addis Ababa, Ethiopia from January to June 2019 (N = 156) Parameters Mean ± SD Median (IQR) Range 95% RI(2.5th-97.5th Percentile Lower limit 95%CI Upper limit 95%CI Absolute CD4 count (Cells/µL) 782.9 ± 216.7 757.5(611.3-925.5) 394–1669 416.9-1218.4 410.7–463.0 1192.6-1263.9 CD4 (%) 44.1 ± 6.5 43.6(39.9–47.3) 22–63 32.1–57.3 27.9–34.9 55.5–62.7 Hemoglobin (g/dL) 14.35 ± 1.264 14.3 (13.4–15.1) 11–18 NA NA NA NA: Not applicable Discussion This study focuses on the values of absolute and percentage CD4cell counts as well as Hbg to determine RI in 156 apparently healthy HIV-negative pregnant women from Addis Ababa, Ethiopia. Pregnancy has a physiological change that leads to the suppression of immunological functions for the survival of the new fetus( 15 )Thus, separate reference intervals for tests that indicate immune status like CD4 count are required during pregnancy to find out the pathological complication of maternal and fetal during the pregnancy period. The study found that the RI CD4cell count was 416.95-1218.4 Cells/µL and 32.1–57.3% in absolute terms, while the mean absolute count was 782.9 and median 757.5 according to Table 3 there is no static difference observed in terms of age, for CD4cell absolute count p value 0.160and for CD4cell% p-value of 0.337. It also the same for Gestational age (Trimester) Even if there is no reference interval was developed in terms of gestation age because of the insufficient sample according to some study findings there is a difference in terms of Gestational age (Trimester)( 16 )study finding was comparable with previous studies done on healthy pregnant women in Ethiopia. ( 6 ) The study also explained that this study finding of the RIs determined in other studies in Ethiopia and elsewhere. There was an inconsistency of RIs from the different countries according to this current study was slightly lower than the study in Cameroonian pregnant women which was 851 Cells/µ( 17 )but comparable with the study finding in India ( 18 )the observed variation maybe because of temporary immune suppression is normally observed during a healthy pregnancy. The hemoglobin RI was not developed because Hemoglobin concentration had a weak negative correlation with gestational age (r=-0.160; p = 0.048) It was difficult to set general RI it needed separation intra gestational age however we didn’t get sufficient sample so we set for this study mean and median. According to this finding some inconsistency was observed with the RI study done on pregnant women in Gojjam, northwest Ethiopia, and Northeast Ethiopia ( 19 , 14 , and 5 ). The hemoglobin mean of this study was 14.35g/dl which is higher than some study done on pregnant women in Africa However; the finding is comparable with other reports 17 ). The observed higher finding may be due to the geographical differences that the study participants from Ethiopia located at a high altitude which is one of the factors affecting hemoglobin value. So study findings indicate that developing of reference intervals is crucial for the good management of fatal and maternal health status. Conclusion Comparing this study's results to those of non-African and African nations, it revealed a decline in both parentage count and absolute CD4+ T cell counts. Therefore, it is essential to establish local reference values for various populations. Limitation The main limitation of this study was the sample size for developing RI interims Gestational age (Trimester) because 120 samples were needed for each according to CLSI. Abbreviations CD4: cluster differentiation, CD4%-cluster differentiation percentage, CD4cluster differentiation absolute count, NCCLS: National committee of clinical laboratory standard, WHO: World Health Organization, Hgb: hemoglobin, RI: reference interval, HIV: human immunodeficiency virus. Declarations The Departmental Research and Ethics Review Committee of Addis Ababa University's Department of Medical Laboratory Sciences gave its approval to the study protocol. Participants in the study were informed about the purpose of the investigation, the risks involved, the benefits of participating, and their right to withdraw at any time. They were also given access to the laboratory results upon request. Availability of data All data that support the findings of this study are available on request from the corresponding author. Disclosure The authors declare that they have no conflict of interest. Author Contributions The authors participate in data collection, analysis, selection of the journal to which the article will be submitted, and are responsible for the final approval of the version to be published . Funding This research was supported by the Ethiopian Public Health Institution (EPHI) and the Ministry of Science and Technology (current MINT) in collaboration with Addis Ababa University. Acknowledgment The authors would like to thank Addis Ababa University, Department of Medical Laboratory Sciences, Ethiopian Public Health Institute and Ministry of Science and Technology (current Ministry of Innovation and Technology) for sponsoring this research. Central Statistical agency, Ministry of Health, Health extension workers, Addis Ababa Health Bureau and institutions which facilitated the data collection process are all gratefully acknowledged for making this study possible. We also thank all study participants who voluntarily participated in this study. References Akinbami AA, Dosunmu AO, Adediran A, Adewunmi AA, Rabiu KA, Osunkalu V. Cluster of differentiation 4 + cell count mean value , reference range and its influencing factors in Human Immunodeficiency Virus-seronegative pregnant women in Lagos.2014;55(2). Zhu J, Yamane H, Paul WE. Differentiation of effector C D4 T cell populations (*). Annu Rev Immunol. 2010; 28:445-89. doi: 10.1146/017;28–34. 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Babker AM, Osama E, Elnaim D. Hematological Changes During All trimesters in Normal Pregnancy. 2020;10(2):1–4. Access O. We are IntechOpen , the world ’ s leading publisher of Open Access books Built by scientists , for scientists TOP 1 %. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; ApprovedGuideline.Clinical and Laboratory standards Institute; 2008 The 2007 population and housing census of Ethiopia: population census commission. Central Statistical Authority. 2012 Angira F, Akoth B, Omolo P, Opollo V, Bornheimer S. Clinical Evaluation of the BD FACSPresto TM Near-Patient CD4 Counter in Kenya. 2016;1–25. PrabhuDas M, Bonney E, Caron K, Dey S, Erlebacher A, Fazleabas A, Fisher S, Golos T, Matzuk M, McCune JM, Mor G, Schulz L, Soares M, Spencer T, Strominger J, Way SS, Yoshinaga K. Immune mechanisms at the maternal-fetal interface: perspectives and challenges. Nat Immunol. 2015 (4):328-34 Malko A, Assegu D, Id F. Hematological and CD 4 + T- cell count reference interval for pregnant women attending antenatal care at Hawassa University Comprehensive Specialized Hospital , Hawassa Southern Ethiopia. 2021;342:1–14. Available from: http://dx.doi.org/10.1371/journal.pone.0249185 Nlinwe NO, Kumenyuy YL, Funwi CP. Establishment of Hematological Reference Values among Healthy Adults in Bamenda , North West Region of Cameroon. 2021;2021. Dayama A, Pandit D, Mudaliar S, Bharadwaj R, Bharucha KE, Shrotri AN, et al. A pilot study on CD4 & CD8 cell counts in healthy HIV seronegative pregnant women. 2003; 198–200. Mulu W, Abera B, Mekonnen Z, Adem Y, Yimer M, Zenebe Y, et al. Haematological and CD4 + T cells reference ranges in healthy adult populations in Gojjam zones in Amhara region , Ethiopia. l. 2010;28:445-89. doi: 10.1146/017;28–34. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 11 Mar, 2025 Read the published version in BMC Research Notes → Version 1 posted Editorial decision: Revision requested 07 Jun, 2024 Editor assigned by journal 05 Jun, 2024 Submission checks completed at journal 05 Jun, 2024 First submitted to journal 30 May, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4503170","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":311706471,"identity":"6ae479df-9f35-4c20-a68a-207e55cff26d","order_by":0,"name":"Dinkenesh 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Institute","correspondingAuthor":false,"prefix":"","firstName":"Ebba","middleName":"","lastName":"Abate","suffix":""},{"id":311706490,"identity":"ca7bf662-47ce-4dd7-947b-2ad159d10274","order_by":11,"name":"Aster Tsegaye","email":"","orcid":"","institution":"Addis Ababa University","correspondingAuthor":false,"prefix":"","firstName":"Aster","middleName":"","lastName":"Tsegaye","suffix":""}],"badges":[],"createdAt":"2024-05-30 12:56:04","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4503170/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4503170/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s13104-025-07171-w","type":"published","date":"2025-03-11T15:57:32+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":59051776,"identity":"ece340fc-5e85-44ce-844a-c56dc3e122ef","added_by":"auto","created_at":"2024-06-25 20:09:30","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":216709,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4503170/v1/5ae9216e72b332d502f8fe3c.png"},{"id":78689146,"identity":"3de74a10-0407-4d00-9fe7-20ada603e59d","added_by":"auto","created_at":"2025-03-17 16:11:53","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":955121,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4503170/v1/d158d8c2-82ef-4696-bd8e-919cdc688d8d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Reference Intervals for CD4 and hemoglobin among apparently healthy pregnant women in Addis Ababa, Ethiopia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eCD4 cells are a subset of T-lymphocytes that act as helper or inducer T-cells. Because they don\u0026rsquo;t kill the pathogen directly instead they induce a signal to other T cells and macrophages to initiate an immune response. The Cell is to mediate adapting immunity for the different pathogens, immunological memory and also participate in autoimmunity, asthma, and allergic responses when during individual become susceptible to a wide range of infectious disorders. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eDuring pregnancy, some physiological changes occur that influence immunological activities that change the CD4 count. It provides a figure of the immune system in health; a higher CD4 count typically indicates the wellness of the immune system condition. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eCD4 count can be affected by different factors starting from sample collection, up to analysis those are pathological, and physiological factors such as the technique and time of blood collection, transport, and storage of the specimens, posture, exercise, methodology, and instrument used to obtain the result (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e) many laboratories lack RI for pregnant women, even though the physiological and biochemical changes in pregnancy influence many of our laboratory tests. Without adequate RI, there is an increased risk of missing significant changes due to pathological conditions and incorrectly interpreting normal changes as pathological events(\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eCertain laboratories have identified this condition and provide RI to pregnant women, even though pregnancy itself causes the same changes in routine laboratory tests. The majority of labs are actually unaware of the typical RI for expectant mothers. The results change depending on the trimester of pregnancy; for instance, the first and last trimesters have the highest hemoglobin concentrations, while the second trimester has the lowest. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e) In comparison to populations in some other African nations and the West, adult Ethiopian populations have reportedly been found to have higher hemoglobin levels. Conversely, during pregnancy, which is marked by numerous changes in practically all organ systems to accommodate the growing and developing fetal placenta, they have a lower CD4 cell count and CD4+/CD8\u0026thinsp;+\u0026thinsp;T-cell ratio (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). Units, leading to a reduction in the majority of laboratory test parameters. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e) According to CLSI recommendation, each laboratory shall establish its reference range because the RI is affected by several factors including ethnicity, nutrition, altitude, geographic location, and pregnancy (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e Therefore, this study was designed to develop CD4 cell count and hemoglobin RI in apparently healthy pregnant women in Addis Ababa which was important as a guide for clinical support and treatment monitoring of patients in disease conditions and clinical trials. The findings will also be used as a reference for future studies.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy design, period, and setting\u003c/h2\u003e \u003cp\u003eFor voluntary, seemingly healthy pregnant women in Addis Ababa, Ethiopia, a community-based cross-sectional study was conducted from January to June 2019.\u003c/p\u003e \u003cp\u003eAddis Ababa lies between 2200 and 2500 meters over ocean level. It was the establishment of distinctive population bunches moreover known as the biggest capital city of the nation. And covers 527 square kilometers of zone in Ethiopia, and it has a 3,384,569 population size concurring to the 2007 census (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e) during the study period, the city was categorized into 10 sub-cities. Based on the population estimate, 4 sub-cities (Yeka, Akaki Kality, Arada, and Kirkos) were chosen.\u003c/p\u003e \u003cp\u003eIt appears that the study included healthy pregnant women in Addis Ababa who were between the ages of 18 and 49 and who satisfied the eligibility requirements. Anemia, chronic renal failure, hypertension, coronary heart disease, diabetes, thyroid disease, liver disease, or any cancer is examples of chronic illnesses. People who use pharmacologically active substances aside from iron and who have received a blood transfusion within the last year are also listed. Additionally, factors like intestinal parasites, malaria, high CRP levels, smoking, drinking, and supplementing with folic acid were disregarded.\u003c/p\u003e \u003cp\u003eThe sampling strategy used in this study was a probability ratio. The sample size was established following the CLSI recommendation that a good way to develop RI is to collect samples from a sufficient number of different individuals to obtain at least 120 samples for analysis. PPS, where size is determined by the number of households in the city's Woredas (previously Kebeles, the smallest administrative units). Due to the size of Addis Abebe, four suburbs Arada, Kirkos, Akaki, and Yeka were chosen for the PPS; as a result, all of the chosen suburban woredas were included. Using Woreda as the sampling frame, systematic random sampling was used to select study participants. Informed consent was obtained before a blood sample was taken for laboratory analysis, and structured questionnaires were utilized to gather socio-demographic information.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSpecimen collection and laboratory analysis\u003c/h2\u003e \u003cp\u003eUsing a vacutainer tube filled with ethylene diamine tetra acetic acid (EDTA) tubes, the sample was collected. Samples were taken prior to 11:00 a.m. to prevent daily variations and are identified with distinct codes. At the National HIV Reference Laboratory of the Public Health Institute of Ethiopia, a point-of-care FACS Presto (BD, San Jose, CA, USA) was used to analyze the CD4 count and hemoglobin in both absolute and percentage terms within eight hours of sample collection. Since 2016, Ethiopia has been using BD FACS PrestoTM CD4 point-of-care technology, which has a contract with FACS Calibur that is acceptable for CD4 cell counts and CD4 (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). The laboratory analyses were conducted strictly in accordance with standard operating procedures in order to guarantee the quality of the results. Furthermore, the external quality system is a part of the National Reference Laboratory.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eSPSS 20 was used to enter and analyze the data. With SPSS 20, the data distribution was evaluated and the data were input and examined. After the distribution of the data was evaluated, the data were compiled, summarized, and shown as absolute numbers with percentages for categorical variables and mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD and medians with IQR values for continuous variables. The nonparametric Spearman correlation test was used to measure correlation, and the nonparametric Manutiny U and Kruskal-Wallis tests were used to compare groups. A non-parametric test was used to calculate RIs, estimating percentages of 2.5, the lower limit, and 97.5, the upper limit, with a 95% confidence interval. The RI was determined to be the 95 percent interval. It was determined that 0.05 was statistically significant.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eEthical Considerations\u003c/h2\u003e \u003cp\u003eRegarding ethical issues, The Helsinki Declaration was followed in the conduct of this study. The Departmental Research and Ethics Review Committee of Addis Ababa University's Department of Medical Laboratory Science examined and approved the study protocol. At the time of enrollment, consent was obtained from voluntary participants, and all participants gave written informed consent. Participants received their CD4\u0026thinsp;+\u0026thinsp;and hemoglobin test results back upon request, utilizing the BD FACSPresto.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSocio-demographic characteristics\u003c/h2\u003e \u003cp\u003e162 apparently healthy pregnant women aged 18\u0026ndash;49 years were included in the determination of CD4 cells and hemoglobin RI. Of these, 6 participants were excluded due to chronic diseases, parasitic infections, HIV, and high CRP. A total of 156 participants participated in the final analysis. The median age of the study participants was 27 (IQR\u0026thinsp;=\u0026thinsp;24\u0026ndash;29) years. More than half (58.33%) of the participants were 25\u0026ndash;34 years old. Most of the participants (88.4%) were married, 37.2% had primary school education and 51.9% were housewives (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSocio-demographic characteristics of study participants in Addis Ababa, Ethiopia from January to June 2019\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eVariables\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentage (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eAge(years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18\u0026ndash;24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e36.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25\u0026ndash;34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e58.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35\u0026ndash;49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eMarital status\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSingle\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMarried\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e138\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e88.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDivorced\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eEducational status\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIlliterate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRead and write\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrimary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e37.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSecondary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e36.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCollege and above\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eOccupation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHousewife\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e51.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGovernment employee\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrivate employee\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e39.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eCD4 + T-cells and Hemoglobin by study participant’s characteristics\u003c/h3\u003e\n\u003cp\u003eThe median value of CD4 count and Hbg concentration by different groups of study participant\u0026rsquo;s characteristics is presented in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Overall status there was no statistically significant difference in CD4 count and hemoglobin concentration with participant\u0026rsquo;s characteristics except hemoglobin concentration in the gestation stage (trimester). The median value of absolute CD4 value was 779 (640\u0026ndash;937) cells/\u0026micro;l 725 (594-927.5)cells/\u0026micro;l and 779(619.5\u0026ndash;916) cells/\u0026micro;l in the first, second, and third trimesters respectively. The hemoglobin concentration of participants in the first trimester (median: 14.9, IQR: 14.3\u0026ndash;15.4) is significantly higher than that of participants in the second (median: 14.1, IQR: 13.3\u0026ndash;14.9) and third ((median: 14.1, IQR: 13.2\u0026ndash;15.0) trimesters (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The hemoglobin concentration of pregnant women is inversely proportional to gestational age.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMedian (IQR) value of CD4 count and hemoglobin concentration by different characteristics of study participants Addis Ababa, Ethiopia from January to June 2019\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"2\"\u003e\n \u003cp\u003eCharacteristics (N\u0026thinsp;=\u0026thinsp;156)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbsoluteCD4 count (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCD4%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHemoglobin (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMedian (IQR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMedian(IQR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMedian(IQR)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eAge(Years)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u0026ndash;24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e682 (594\u0026ndash;875)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e42.6 (39.4\u0026ndash;46.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.3(13.2\u0026ndash;15.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u0026ndash;34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e794 (621.5\u0026ndash;961)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.2(40.1\u0026ndash;48.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.3(13.5\u0026ndash;15.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35\u0026ndash;49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e730 (590\u0026ndash;925)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e41.8(38.1\u0026ndash;44.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.2(13.4\u0026ndash;15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eGestation stages (Trimester)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1st (n\u0026thinsp;=\u0026thinsp;33)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e779 (640\u0026ndash;937)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.2(39.0-47.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e*14.9(14.3\u0026ndash;15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2nd (n\u0026thinsp;=\u0026thinsp;49)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e725(594-927.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.4(40.2\u0026ndash;48.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.1(13.3\u0026ndash;14.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3rd (n\u0026thinsp;=\u0026thinsp;74)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e779(619.5\u0026ndash;916)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.8(40.0-46.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.1(13.2\u0026ndash;15.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\n \u003cp\u003eEducation status\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIlliterate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e726 (606\u0026ndash;859)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.9(41.1\u0026ndash;46.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.7(13.6\u0026ndash;15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eRead and write\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e931(750.5-1101.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e46(37.0-47.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.3(14.6\u0026ndash;16.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePrimary\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e791.5(619.0-934.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44(40.1\u0026ndash;47.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.4(13.7\u0026ndash;15.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSecondary\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e723(565.5\u0026ndash;899.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e42.5(39.1\u0026ndash;47.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1(13.2\u0026ndash;14.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCollege and above\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e804.5(719.3-922.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.3(41.3\u0026ndash;50.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.5(13.2\u0026ndash;15.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eOccupational status\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHousewife\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e750.5(608\u0026ndash;930)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.5(40.2\u0026ndash;46.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.3(13.5\u0026ndash;15.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGovernment employee\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e804.5(701\u0026ndash;1029)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.2(40.2\u0026ndash;50.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.7(13.4\u0026ndash;15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePrivate employee\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e761(601\u0026ndash;911)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.8(39.3\u0026ndash;48.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.3(13.3\u0026ndash;15.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eBMI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUnderweight\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e731.5(629.3-867.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37.2(36.0-52.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13.1(12.9\u0026ndash;14.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHealthy Weight\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e736.5(596.3\u0026ndash;897)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.5(40.1\u0026ndash;47.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.2(13.4\u0026ndash;15.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOverweight\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e775(627.8\u0026ndash;983)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.4(39.8\u0026ndash;46.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.5(13.6\u0026ndash;15.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eObesity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e918(705.5\u0026ndash;1118)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.9(41.8\u0026ndash;53.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.0(13.9\u0026ndash;15.6)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eFolate supplementation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSupplemented\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e768.5(594.8-929.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.1(39.7\u0026ndash;46.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.0(13.2\u0026ndash;14.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Supplemented\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e746.5(615.0-926.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.2(40.0-48.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.4(13.5\u0026ndash;15.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eIron supplementation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSupplemented\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e768.5(613.0-925.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.7(40.2\u0026ndash;47.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.3(13.4\u0026ndash;15.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Supplemented\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e719.5(565.8-927.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.0(37.8\u0026ndash;46.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.8(13.0-15.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e*p\u0026thinsp;=\u0026thinsp;0.018\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003eCorrelation of CD4 count and hemoglobin concentration with gestational age\u003c/h2\u003e\n \u003cp\u003eWe assessed the correlation of absoluteCD4 count, CD4% and hemoglobin concentration with gestational age (Fig.\u0026nbsp;1). No significant correlation with gestational age for both CD4 absolute count (r=-0.042; p\u0026thinsp;=\u0026thinsp;0.605) and CD4% (r=-0.006; p\u0026thinsp;=\u0026thinsp;0.944) observed, while Hemoglobin concentration had weak negative correlation with gestational age (r=-0.160; p\u0026thinsp;=\u0026thinsp;0.048).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003e\u003cstrong\u003eFigure1\u003c/strong\u003e\u003c/h2\u003e\n \u003cp\u003e- Correlation of CD4 absolute count (cells/\u0026micro;L), CD4% and hemoglobin concentration (g/dL) with gestational age (week) in pregnant women Correlation of A) absoluteCD4 count Vs gestational age, B) CD4% Vs gestational age and C hemoglobin Vs gestational age. Non-parametric spearman correlation test used to assess correlation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eAbsolute and Percentage CD4cells and Hemoglobin Reference Intervals\u003c/h2\u003e\n \u003cp\u003eAbsoluteCD4 count ranged between 394\u0026ndash;1699 cells/\u0026micro;L while CD4% ranged 22\u0026ndash;63% and Hgb 11\u0026ndash;18 g/dL. According to Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, the ranges for CD4% and absolute CD4\u0026thinsp;+\u0026thinsp;T-cell counts were 32.1\u0026ndash;57.3% and 416.9\u0026ndash;1218.4 cells/\u0026micro;L, respectively.\u003c/p\u003e\n \u003cdiv\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMean, Median, Range and 95% reference interval of CD4 and hemoglobin for healthy pregnant women in Addis Ababa, Ethiopia from January to June 2019 (N\u0026thinsp;=\u0026thinsp;156)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMedian (IQR)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eRange\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e95% RI(2.5th-97.5th Percentile\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLower limit 95%CI\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eUpper limit 95%CI\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbsolute CD4 count (Cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e782.9\u0026thinsp;\u0026plusmn;\u0026thinsp;216.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e757.5(611.3-925.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e394\u0026ndash;1669\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e416.9-1218.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e410.7\u0026ndash;463.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1192.6-1263.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCD4 (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.1\u0026thinsp;\u0026plusmn;\u0026thinsp;6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43.6(39.9\u0026ndash;47.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u0026ndash;63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.1\u0026ndash;57.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e27.9\u0026ndash;34.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55.5\u0026ndash;62.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHemoglobin (g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.35\u0026thinsp;\u0026plusmn;\u0026thinsp;1.264\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.3 (13.4\u0026ndash;15.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u0026ndash;18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003eNA: Not applicable\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study focuses on the values of absolute and percentage CD4cell counts as well as Hbg to determine RI in 156 apparently healthy HIV-negative pregnant women from Addis Ababa, Ethiopia. Pregnancy has a physiological change that leads to the suppression of immunological functions for the survival of the new fetus(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e)Thus, separate reference intervals for tests that indicate immune status like CD4 count are required during pregnancy to find out the pathological complication of maternal and fetal during the pregnancy period.\u003c/p\u003e \u003cp\u003eThe study found that the RI CD4cell count was 416.95-1218.4 Cells/µL and 32.1–57.3% in absolute terms, while the mean absolute count was 782.9 and median 757.5 according to Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e there is no static difference observed in terms of age, for CD4cell absolute count p value 0.160and for CD4cell% p-value of 0.337. It also the same for Gestational age (Trimester) Even if there is no reference interval was developed in terms of gestation age because of the insufficient sample according to some study findings there is a difference in terms of Gestational age (Trimester)(\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e)study finding was comparable with previous studies done on healthy pregnant women in Ethiopia. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eThe study also explained that this study finding of the RIs determined in other studies in Ethiopia and elsewhere. There was an inconsistency of RIs from the different countries according to this current study was slightly lower than the study in Cameroonian pregnant women which was 851 Cells/µ(\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e)but comparable with the study finding in India (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e)the observed variation maybe because of temporary immune suppression is normally observed during a healthy pregnancy.\u003c/p\u003e \u003cp\u003eThe hemoglobin RI was not developed because Hemoglobin concentration had a weak negative correlation with gestational age (r=-0.160; p = 0.048) It was difficult to set general RI it needed separation intra gestational age however we didn’t get sufficient sample so we set for this study mean and median.\u003c/p\u003e \u003cp\u003eAccording to this finding some inconsistency was observed with the RI study done on pregnant women in Gojjam, northwest Ethiopia, and Northeast Ethiopia (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, and \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). The hemoglobin mean of this study was 14.35g/dl which is higher than some study done on pregnant women in Africa However; the finding is comparable with other reports \u003cb\u003e17\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eThe observed higher finding may be due to the geographical differences that the study participants from Ethiopia located at a high altitude which is one of the factors affecting hemoglobin value. So study findings indicate that developing of reference intervals is crucial for the good management of fatal and maternal health status.\u003c/p\u003e "},{"header":"Conclusion","content":"\u003cp\u003eComparing this study\u0026apos;s results to those of non-African and African nations, it revealed a decline in both parentage count and absolute CD4+ T cell counts. Therefore, it is essential to establish local reference values for various populations.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLimitation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe main limitation of this study was the sample size for developing RI interims Gestational age (Trimester) because 120 samples were needed for each according to CLSI.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eCD4: cluster differentiation, CD4%-cluster differentiation percentage, CD4cluster differentiation absolute count, NCCLS: National committee of clinical laboratory standard, WHO: World Health Organization, Hgb: hemoglobin, RI: reference interval, HIV: human immunodeficiency virus.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe Departmental Research and Ethics Review Committee of Addis Ababa University\u0026apos;s Department of Medical Laboratory Sciences gave its approval to the study protocol. Participants in the study were informed about the purpose of the investigation, the risks involved, the benefits of participating, and their right to withdraw at any time. They were also given access to the laboratory results upon request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data that support the findings of this study are available on request from the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eDisclosure\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors participate in data collection, analysis, selection of the journal to which the article will be submitted, and are responsible for the final approval of the version to be published\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the Ethiopian Public Health Institution (EPHI) and the Ministry of Science and Technology (current MINT) in collaboration with Addis Ababa University.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank Addis Ababa University, Department of Medical Laboratory Sciences, Ethiopian Public Health Institute and Ministry of Science and Technology (current Ministry of Innovation and Technology)\u0026nbsp;for sponsoring this research. Central Statistical agency, Ministry of Health, Health extension workers, Addis Ababa Health Bureau and institutions which facilitated the data collection process are all gratefully acknowledged for making this study possible. We also thank all study participants who voluntarily participated in this study.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAkinbami AA, Dosunmu AO, Adediran A, Adewunmi AA, Rabiu KA, Osunkalu V. Cluster of differentiation 4 + cell count mean value , reference range and its influencing factors in Human Immunodeficiency Virus-seronegative pregnant women in Lagos.2014;55(2).\u003c/li\u003e\n\u003cli\u003eZhu J, Yamane H, Paul WE. Differentiation of effector C D4 T cell populations (*). Annu Rev Immunol. 2010; 28:445-89. doi: 10.1146/017;28\u0026ndash;34.\u003c/li\u003e\n\u003cli\u003eMair C, Hawes SE, Agne HD. Factors associated with CD 4 lymphocyte counts in HIV-negative Senegalese individuals. 2007;432\u0026ndash;40. \u003c/li\u003e\n\u003cli\u003eUche EI. Factors influencing CD4 cell count in HIV-positive pregnant women in a secondary health center in Lagos , Nigeria. 2015;115\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eWomen P, Buea IN. REFERENCE VALUES OF CD4-LYMPHOCYTE COUNTS IN HIV SERONEGATIVE. 2012;13(1):28\u0026ndash;33. \u003c/li\u003e\n\u003cli\u003eId MF, Id MM, Ebrahim E, Demsiss W, Tilahun M, Id AT. Common hematological parameters reference intervals for apparently healthy pregnant and non-pregnant women of South Wollo Zone , Amhara Regional State ,. 2022;1\u0026ndash;20. Available from: http://dx.doi.org/10.1371/journal.pone.0270685\u003c/li\u003e\n\u003cli\u003eLarsson A, Palm M, Hansson LO, Axelsson O. Reference values for clinical chemistry tests during normal pregnancy. BJOG. 2008 Jun;115(7):874-81\u003c/li\u003e\n\u003cli\u003eGreer LG. A Reference Table for Clinicians. 2009;114(6):1326\u0026ndash;31. \u003c/li\u003e\n\u003cli\u003ePress D. Immunological and hematological reference intervals among HIV-seronegative pregnant women in northwest Ethiopia. 2017;145\u0026ndash;50. \u003c/li\u003e\n\u003cli\u003eBabker AM, Osama E, Elnaim D. Hematological Changes During All trimesters in Normal Pregnancy. 2020;10(2):1\u0026ndash;4. \u003c/li\u003e\n\u003cli\u003eAccess O. We are IntechOpen , the world \u0026rsquo; s leading publisher of Open Access books Built by scientists , for scientists TOP 1 %. \u003c/li\u003e\n\u003cli\u003eDefining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; ApprovedGuideline.Clinical and Laboratory standards Institute; 2008\u003c/li\u003e\n\u003cli\u003eThe 2007 population and housing census of Ethiopia: population census commission. Central Statistical Authority. 2012\u003c/li\u003e\n\u003cli\u003eAngira F, Akoth B, Omolo P, Opollo V, Bornheimer S. Clinical Evaluation of the BD FACSPresto \u003csup\u003eTM\u003c/sup\u003e Near-Patient CD4 Counter in Kenya. 2016;1\u0026ndash;25. \u003c/li\u003e\n\u003cli\u003ePrabhuDas M, Bonney E, Caron K, Dey S, Erlebacher A, Fazleabas A, Fisher S, Golos T, Matzuk M, McCune JM, Mor G, Schulz L, Soares M, Spencer T, Strominger J, Way SS, Yoshinaga K. Immune mechanisms at the maternal-fetal interface: perspectives and challenges. Nat Immunol. 2015 (4):328-34\u003c/li\u003e\n\u003cli\u003eMalko A, Assegu D, Id F. Hematological and CD 4 + T- cell count reference interval for pregnant women attending antenatal care at Hawassa University Comprehensive Specialized Hospital , Hawassa Southern Ethiopia. 2021;342:1\u0026ndash;14. Available from: http://dx.doi.org/10.1371/journal.pone.0249185\u003c/li\u003e\n\u003cli\u003eNlinwe NO, Kumenyuy YL, Funwi CP. Establishment of Hematological Reference Values among Healthy Adults in Bamenda , North West Region of Cameroon. 2021;2021. \u003c/li\u003e\n\u003cli\u003eDayama A, Pandit D, Mudaliar S, Bharadwaj R, Bharucha KE, Shrotri AN, et al. A pilot study on CD4 \u0026amp; CD8 cell counts in healthy HIV seronegative pregnant women. 2003; 198\u0026ndash;200. \u003c/li\u003e\n\u003cli\u003eMulu W, Abera B, Mekonnen Z, Adem Y, Yimer M, Zenebe Y, et al. Haematological and CD4 + T cells reference ranges in healthy adult populations in Gojjam zones in Amhara region , Ethiopia. l. 2010;28:445-89. doi: 10.1146/017;28\u0026ndash;34. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"CD4 count, %CD4, Hemoglobin, Reference Interval, Ethiopia","lastPublishedDoi":"10.21203/rs.3.rs-4503170/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4503170/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cb\u003eBackground\u003c/b\u003e, Most African countries, including Ethiopia, have not developed local well-defined reference intervals for immuno-hematological testes interims of pregnant women, so we were forced to use reference intervals from manufacturer insert kit values derived from non-Africans. This is not appropriate because CD4 cell count tests are affected by ethnic and environmental factors Therefore, this study aimed to develop RI for CD4 cell count and hemoglobin levels for apparently healthy pregnant women in Addis Ababa, Ethiopia.\u003c/p\u003e \u003cp\u003e\u003cb\u003eMethod\u003c/b\u003e From January to June 2019, 162 pregnant women who appeared healthy were enrolled in this community-based cross-sectional study. Social and medical data were gathered through a structured questionnaire following the acquisition of informed consent. BD FACS Presto, a near-patient CD4 counter, was used to obtain the CD4 count and hemoglobin (Hgb) levels from whole blood using a nonparametric test that estimated 2.5th (lower limit) and 97.5th (upper limit) percentiles with 95% CIs. Data were entered and analyzed using SPSS 20 and P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003cp\u003e \u003cb\u003eResults\u003c/b\u003e: After excluding six pregnant women who did not pass the screening tests, 156 apparently healthy pregnant women participants who were 18\u0026ndash;49 years old were included in the final analysis. The majority of them were in the 2nd trimester, from the total participants (51.9%) were housewives, and (37.2. %) attained at least primary-level education. The medians of CD4%-and CD4# counts with inter-quartile ranges [IQR] were 757.5 [611.3-925.5] cells/\u0026micro;Land 43.62% [39.9\u0026ndash;47.3], respectively while the median and IQR. Hemoglobin values were 14.3gm/dL [13.40\u0026ndash;15.10].\u003c/p\u003e \u003cp\u003eThe absolute CD4 cell count reference intervals were 416.9-1218.4 cells/\u0026micro;L, %CD4\u0026thinsp;+\u0026thinsp;32.1\u0026ndash;57.3%, there is no significant difference between the trimester and age difference was observed.\u003c/p\u003e \u003cp\u003e \u003cb\u003eConclusion\u003c/b\u003e: Comparing this study's results to those of non-African and African nations, it revealed a decline in both parentage count and absolute CD4\u0026thinsp;+\u0026thinsp;T cell counts. Therefore, it is essential to establish local reference values for various populations.\u003c/p\u003e","manuscriptTitle":"Reference Intervals for CD4 and hemoglobin among apparently healthy pregnant women in Addis Ababa, Ethiopia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-25 20:09:25","doi":"10.21203/rs.3.rs-4503170/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-06-07T09:23:30+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-06-05T08:03:26+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-06-05T08:03:16+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Research Notes","date":"2024-05-30T12:54:48+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-research-notes","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"resn","sideBox":"Learn more about [BMC Research Notes](http://bmcresnotes.biomedcentral.com)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/resn/default.aspx","title":"BMC Research Notes","twitterHandle":"@BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"71c186ec-095b-45ba-b7b9-3f3da552925a","owner":[],"postedDate":"June 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-03-17T16:06:39+00:00","versionOfRecord":{"articleIdentity":"rs-4503170","link":"https://doi.org/10.1186/s13104-025-07171-w","journal":{"identity":"bmc-research-notes","isVorOnly":false,"title":"BMC Research Notes"},"publishedOn":"2025-03-11 15:57:32","publishedOnDateReadable":"March 11th, 2025"},"versionCreatedAt":"2024-06-25 20:09:25","video":"","vorDoi":"10.1186/s13104-025-07171-w","vorDoiUrl":"https://doi.org/10.1186/s13104-025-07171-w","workflowStages":[]},"version":"v1","identity":"rs-4503170","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4503170","identity":"rs-4503170","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}