Development of serum lipoprotein (a) detection using latex enhanced immunoturbidimetry

Development of serum lipoprotein (a) detection using latex enhanced immunoturbidimetry | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Development of serum lipoprotein (a) detection using latex enhanced immunoturbidimetry Yanyan Liu, Meijiao Li, Hao Zhang, Le Gao, Jitao Liu, Yue Hou, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4549466/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Mar, 2025 Read the published version in Biotechnology Letters → Version 1 posted 5 You are reading this latest preprint version Abstract Background Lipoprotein (a) (Lp (a)) is indeed a significant factor in cardiovascular health, as it is a product of low-density lipoprotein cholesterol-like particles that bind to apolipoprotein (a). Elevated levels of Lp (a) have been linked to an increased risk of cardiovascular diseases (CVD), hastening disease progression and raising CVD mortality rates. However, the absence of standardized measurement methods for Lp (a) contributes to diagnostic uncertainties. Method A quantitative measurement method for serum Lp (a) was developed using fully automated latex-enhanced particle immunoturbidimetry technology represents a significant advancement in diagnostic capabilities. The key parameters such as repeatability, stability, linearity, and method comparison were evaluated to ensure the accuracy of the assay. Result The Lp (a) in samples was recognized by carboxylated latex particles covalently coated with anti-Lp (a) antibodies. The content of Lp (a) was quantified by measuring the changes in turbidity generated by agglutination at 600 nm. With precision CV% within the batch of 1.10% and inter-batch precision CV% of 1.79%, it demonstrates reliable performance using Randox biochemical quality control samples. The detection limit of 7 mg/L and a high correlation coefficient (R 2 = 0.9946) at concentrations of 0-1500 mg/L further validate its effectiveness. Conclusion The quantitative determination method of serum Lp (a) based on latex-enhanced immunoturbidimetric analysis indeed provides rapid results, high accuracy, and automation, making it suitable for routine clinical testing. This method relies on the interaction between Lp (a) and latex particles, allowing for efficient measurement in serum samples. Cardiovascular disease Lipoprotein (a) Measurement value Atherosclerosis Latex enhanced turbidimetric immunoassay Abnormal blood lipids Figures Figure 1 Figure 2 Introduction Atherosclerosis is the root cause of most cardiovascular diseases (CVD), which causes more than 17.9 million deaths every year (Murray, 2022 ). In recent years, although significant progress has been made in the prevention and treatment of CVD, it is still the main cause of incidence rate and mortality (Bułdak, 2022 ; Mensah, Roth, & Fuster, 2019 ; Timmis et al., 2018 ; Xu, Murphy, Kochanek, & Arias, 2022 ; Zhao, Liu, Wang, Zhang, & Zhou, 2019 ). The occurrence of CVD is related to many factors, such as obesity, type 2 diabetes (T2DM) and metabolic syndrome (Mensah et al., 2019 ). In addition, old age, hypertension, dyslipidemia, smoking and lack of exercise are traditional risk factors leading to accelerated atherosclerosis and early cardiovascular events (Yang et al., 2023 ). Genetic susceptibility is a key factor in the risk and progression of CVD. Although many standardized management methods have been used to address CVD risk factors, there are still significant CVD risks in some populations. Elevated levels of lipoprotein (a) (Lp (a)) in plasma are an independent and primarily gene determined pathogenic risk factor for CVD (Duarte Lau & Giugliano, 2022 ; Ward et al., 2021 ). In 1963, K á re Berg first demonstrated that Lp (a) is a unique lipoprotein similar to low-density lipoprotein (LDL) (Berg, 1963 ). Lp (a) can accumulate in the subendothelial space and has the potential to cause thrombosis and atherosclerosis (Rath et al., 1989 ). Recent experiments have shown that high levels of Lp (a) lead to a higher incidence of CVD, accelerate disease progression, and increase the mortality rate of CV (Kamstrup, Benn, Tybjaerg-Hansen, & Nordestgaard, 2008 ; Lampsas et al., 2022 ; Ruscica, Sirtori, Corsini, Watts, & Sahebkar, 2021 ). In addition, due to the lack of standardized Lp (a) measurement methods, there is diagnostic uncertainty in this field (Lampsas et al., 2023 ). Latex enhanced turbometric immunoassay (LETIA) is an immunoassay technique that uses microparticles as markers. The core principle of this method is based on antigen antibody reactions, using latex coated with antibodies to react with specific analytes. Trigger the relationship between particle aggregation and analyte concentration through fast and simple measurement methods such as turbidimetry (Duan et al., 2023 ; Xia et al., 2017 ). In this method, the carrier particles used are polystyrene latex microspheres of varying diameters (Xia et al., 2017 ), which can bind to protein molecules through appropriate coupling methods. The content of the substance to be detected in the sample can be determined by detecting the degree of reduction in the transmittance of the solution through a spectral analyzer at a specific wavelength (Conde-Sánchez, Roldán-Fontana, Chueca-Porcuna, Pardo, & Porras-Gracia, 2010 ; Dong et al., 2020 ; Duan et al., 2023 ; Favresse et al., 2018 ; Lapić et al., 2020 ). Latex enhanced immunoturbidimetric analysis has highly automated characteristics and is suitable for large-scale clinical sample detection. The objective of this study is to establish an Lp (a) utilizing latex-enhanced immunoturbidimetric analysis, adhering to guidelines set forth by the Clinical and Laboratory Standards Institute (CLSI) (Plebani & Lippi, 2023 ). This reagent kit offers the benefits of automated analysis, precision, rapidity, and resilience to interference. Below, we outline the characteristics and performance of this method, essential for validating its efficacy and establishing its reliability in clinical applications. Materials and Methods Instrument All measurements were performed with Hitachi 7100 fully automated biochemical analyzer (HITACHI, Japan). Samples The samples used in this study were artificial serum matrix (Braveds Biotechnology Co., Ltd., Shenzhen, China) and biochemical quality control samples (2833CH, 2835CH, Randox Company, UK) with different concentrations of recombinant antigens added (China, Yuansheng Biotechnology (Shanghai) Co., Ltd.) for this study, and there is no risk of clinical sample use. Lp (a) antibody The rabbit polyclonal antibody targeting human Lp (a) is produced by Jianzhong Biotechnology Co., Ltd. (Ningbo, China). The antibody exhibited no cross-reactivity with apolipoprotein B and fibrinogen and was stored at 4℃. Calibration products Lp (a) antigen (China, Yuansheng Biotechnology (Shanghai) Co., Ltd., concentration 2000mg/L) was used as a calibration sample, diluted with purified water to prepare standard solutions with concentrations of 0, 300, 600, 900, 1200, and 1500mg/L. In addition, Randox Lp (a) quality control products (batch numbers 2833CH, 2835CH, UK, Randox company) were used as secondary standards. The concentration of Lp (a) (295mg/L, 446mg/L) was repeated using latex enhanced immunoturbidimetric analysis (5 days, n = 10 per day). The calibration sample was stored in a plastic bottle at -30℃ or freeze-dried under 600 mmol/L sucrose conditions (Borque, Maside, Rus, & del Cura, 1993 ). Latex reagents 10% of solid carboxylated polystyrene latex particles with a diameter of 95nm (JSR-P0011), purchased from JSR Corporation in Japan, were used. Following the method described in the literature, covalent coupling was performed between the Lp (a) antibody and JSR-P0011 latex particles (Borque et al., 1989 ). The obtained 0.5% solid coated particle solution was stored at 4℃. The original solution should be frozen or freeze-dried for long-term storage. Measurement procedure On the Hitachi 7100 fully automatic biochemical analyzer, the quantitative addition of samples and reagents was meticulously carried out. A comprehensive 6-point calibration curve was meticulously constructed, spanning concentrations from 0 to 1500 mg/L, with each concentration meticulously tested thrice for precision. The procedural steps are outlined as follows: Commence by dispensing 200 µL of glycine buffer (comprising 0.1 mol/L glycine, 0.15 mol/L NaCl, 10 g/L bovine serum albumin, 30 g/L polyethylene glycol, and 0.6 mmol/L sodium azide, pH adjusted to 8.2). Subsequently, introduce 50 µL of latex reagent into the buffer to initiate the reaction. Follow by adding 3 µL of the experimental sample to the reaction mixture. Proceed to monitor the ensuing reaction at a wavelength of 600 nm over a duration of 5 minutes, meticulously maintaining the temperature at 37°C. Comparison with commercially available reagent kits Compare the performance of the Lp (a) immunolatex reagent with similar products available in the market (Maccure, China, Sichuan Mike Biotechnology Co., Ltd.) by conducting linear fitting and scatter plot analyses on the test results obtained from 50 serum samples within the testing range. Interference substances The interference study followed the Glick program (Glick, Ryder, & Jackson, 1986 ), adding different concentrations of antigens to the artificial serum matrix to prepare basic samples. By adding ascorbic acid (200, 400, 800, 1000, 2000 mg/L, China, Ningbo National Pharmaceutical Group Chemical Reagent Co., Ltd.), fat emulsion (0.25, 0.50, 1.00, 1.50, 2.00%, China, Anhui Fengyuan Pharmaceutical Co., Ltd.), bilirubin (100, 200, 400, 600, 800 µ Mol/L, China, Shanghai Alfa Aesar (China) Chemical Co., Ltd.) and hemoglobin (0.5, 1.0, 2.0, 4.0, 6.0g/L, China, Ningbo National Pharmaceutical Group Chemical Reagent Co., Ltd.) were added to the base sample. Each sample was tested three times, and the average value was taken to calculate the interference rate and evaluate its impact on the measurement results. The interference rate is calculated as the interference concentration divided by the base sample concentration, multiplied by 100%. An interference rate within the range of ± 10% indicates no significant interference, while a rate exceeding ± 10% signifies significant interference. Statistics To investigate the relationship between latex-enhanced immunoassay and commercially available test kits, we will undertake Passing-Bablok regression and Bland-Altman analysis (Fu et al., 2024 ). These methods will help assess the agreement and bias between the two measurement techniques. Additionally, we will calculate the correlation coefficient of Lp (a) values using linear regression analysis. Furthermore, samples at three concentration levels will be utilized to evaluate deviation, coefficient of variation (CV), and recovery rate, providing insights into accuracy, precision, and recovery efficiency across different concentrations. Results Calibration curve Lp (a) measurement was conducted using a 6-point calibration program on the Hitachi 7100 fully automated biochemical analyzer. The analysis range extends from 0 to 1500 mg/L (Fig. 1 ). The linear equation demonstrates a strong linear relationship between each concentration and the detection results, encompassing both normal and pathological serum values. In case the concentration exceeds 1500 mg/L, physiological saline was diluted fourfold and subsequently analyzed automatically by the instrument. The stability of the calibration curve is guaranteed for up to 30 days when reagents are stored at 2 to 8°C. Inaccuracy Conduct 20 consecutive measurements of the Randox quality control substance (2833CH) and observe the imprecision density (Table 1 ). The coefficient of variation within the batch is 1.10%, indicating excellent repeatability in measurement results. Furthermore, when measuring the same Randox quality control sample (2835CH) over 20 consecutive days, the inter-batch coefficient of variation was 1.79%, demonstrating the immunolatex turbidity method's accuracy (Fu et al., 2024 ). Table 1 Results of imprecision measurement for Lp (a) immune latex Project Name Randox Quality Control (2833CH) Randox Quality Control (2835CH) Measurement frequency 20 20 Average value (‾χ) 304.2 464.2 Maximum value ( X max ) 306.3 476.7 Minimum value ( X min ) 303.0 453.0 Standard deviation (SD) ( SD ) 3.34 8.31 Coefficient of variation ( CV ) 1.10% 1.79% Sensitivity To assess the linear relationship, continuously dilute two distinct Randox quality control samples (2833CH, 2835CH). The linear regression coefficient (R 2 ) between Lp (a) and dilution for all samples is 0.994. Within the measurement range, the deviation between the measured value and the theoretical value must not exceed 5%. To determine the lower detection limit, calculate the average value of 20 repeated measurements of physiological saline solution (zero signal) and add 3 standard deviations, resulting in a lower detection limit of 7mg/L. Interference substances Interference samples of varying concentrations are measured with the sample concentration serving as the reference value. When the sample's ascorbic acid concentration is ≤ 2000 mg/L, fat emulsion concentration is ≤ 2.00%, bilirubin concentration is ≤ 800 µmol/L, and hemoglobin concentration are ≤ 6g/L, the interference rate of interferents on Lp (a) determination should be ≤ ± 10% (refer to Table 2 ). This demonstrates the absence of endogenous interference in the determination of the immunolatex reagent. Table 2 Distribution of interference rates (%) of four substances ★ Ascorbic acid (mg/L) Fat emulsion (%) Bilirubin (µmol/L) Hemoglobin (g/L) A B A B A B A B 200 -2.5 0.25 -1.5 100 + 2.2 0.5 -2.2 400 + 0.5 0.5 -0.7 200 + 4.0 1.0 -4.3 800 -6.2 1.0 -2.6 400 + 2.8 2.0 -3.0 1000 -7.5 1.5 -3.1 600 + 5.1 4.0 -7.5 2000 -8.4 2.0 -5.2 800 + 4.9 6.0 -9.3 ★ A: Concentration; B: Interferencerate Recovery rate of spiking Randox quality control (2835CH) with known concentration was added to 5 mixed serum samples and measured. The results of the spiked recovery rate are shown in Table 3 , with a range of 96.5% -105.9%. The average recovery rate is 100.98%. Table 3 Recovery rate of Lp (a) immunolatex determination Heoretical spiked value (mg/L) Measured value (mg/L) Recovery rate (%) 1 129.1 125.6 97.3 2 171.2 182.8 105.9 3 361.9 354.1 97.8 4 439.5 453.0 103.1 5 538.2 519.7 96.5 Correlation coefficient Using the Hitachi 7100 automatic biochemical analyzer, 50 serum samples were simultaneously tested with the Lp (a) immunolatex assay reagent and the control Lp (a) immunolatex reagent (from Maccure). The method comparison results are displayed in Fig. 2 The serum Lp (a) values ranged from 9.1 to 1041.1 mg/L. There was a strong correlation observed between the immune latex reagent and the Maccure reagent samples, with no significant difference noted ( p < 0.01). Discussion Research shows that Lp (a) is an independent risk factor for atherosclerosis and thrombosis (Bułdak, 2022 ; Mensah et al., 2019 ; Timmis et al., 2018 ; Xu et al., 2022 ; Yang et al., 2023 ). However, currently there are methodological differences in Lp (a) detection in clinical laboratories due to issues with methods and standardization. Traditional Lp (a) measurement methods typically involve cumbersome manual analysis techniques. These methods generally suffer from issues such as unstable precision and long detection time. In order to overcome these problems, several automatic immune turbidity analysis methods have been developed in recent years (Borque, Rus, del Cura, Maside, & Escanero, 1993 ; Tiran, Tiran, Hojas, Kostner, & Wilders-Truschnig, 1993 ), but there are significant differences in the detection results between these methods (Borque, Maside, & Iglesias, 1993 ). One of the main issues in detecting serum Lp (a) in the past was the difference in the purity of Lp (a) antibodies, as well as the difference in particle size selection due to carboxylated polystyrene microspheres (Tiran et al., 1993 ). This study utilizes carboxylated latex particles, 95 nm in diameter, to covalently coat Lp (a) antibodies. The sample and coated latex particles underwent a 5-minute incubation at 37°C, followed by measuring turbidity change at 600 nm to quantify the agglutination reaction. Optimal detection efficacy is achieved with a coupling ratio of 5:1 between latex microspheres and Lp (a) antibody, resulting in larger signal generation due to the smaller volume of microspheres. This analysis method offers rapid, accurate, and fully automated measurements on the Hitachi 7100 automatic biochemical analyzer. Using Randox biochemical quality control samples, precision evaluation revealed an intra-batch precision CV% of 1.10% and an inter-batch precision CV% of 1.79%. The detection limit (LoD) for Lp (a) is 7 mg/L, with a correlation coefficient R 2 = 0.9946 within the 0-1500 mg/L concentration range. Notably, interference from bilirubin, fat emulsion, hemoglobin, and ascorbic acid is absent. When compared with a commercially available latex-enhanced particle immunoturbidimetric Lp (a) assay reagent, the developed reagent exhibited a strong correlation (R 2 = 0.9972) on the Hitachi 7100 analyzer. Overall, this method demonstrates superior comprehensive performance compared to previously reported Lp (a) measurement methods. In conclusion, our study confirms the viability of latex-enhanced particle immunoturbidimetry for serum Lp (a) determination on the Hitachi 7100 fully automated biochemical analyzer. This method demonstrates speed, excellent linearity, precision, and full automation. Through rigorous performance verifications, the particle enhancement method meets expected criteria, thus proving its potential for widespread application in clinical and related fields for Lp (a) detection. Declarations Author Contributions Conceptualization: YY L, Y H and JC X; Data curation: YY L, MJ L, Y H and JT L; Formal analysis: YY L, L G, Y H and JC X; Funding acquisition: H Z, Y H and JC X; Methodology:YY L, H Z, L G and Y H; Visualization: YY L, MJ L and JT L; Writing original draft: YY L, MJ L and JT L; Writing - review &editing: YY L, H Z, L G ,JC X and Y H; Supervision: Y H and JC X. Acknowledgments The authors have no potential conflicts of interest. Special thanks to Dr. Lipeng Xu from Jilin Getein Biotechnology Co., Ltd. for his valuable assistance during the experiment. This work was supported by Science and Technology Department of Jilin Province (Grant No.20220401085YY). Confict of interest The authors have no relevant fnancial or non-fnancial interests to disclose. Ethical approval This article does not contain any studies with human participants or animals performed by any of the authors. Consent to participate This article does not contain any studies with human participants or animals performed by any of the authors. Consent to publish This article does not contain any studies with human participants or animals performed by any of the authors Data availability The data generated in this study are available upon request from the corresponding author. Conflict of interest The authors declare that there are no confict of interests. References Berg, K. (1963). A NEW SERUM TYPE SYSTEM IN MAN--THE LP SYSTEM. Acta Pathol Microbiol Scand , 59, 369-382. doi:10.1111/j.1699-0463.1963.tb01808.x Borque, L., Cambiaso, C., Lavenne, E., Leautaud, P., Mareschal, J. C., & Collet-Cassart, D. (1989). Immunoassay by particle counting for coagulation testing: application to the determination of antithrombin III, von Willebrand factor antigen (vWF:Ag) and plasminogen. J Clin Chem Clin Biochem , 27(3), 175-179. doi:10.1515/cclm.1989.27.3.175 Borque, L., Maside, C., & Iglesias, A. (1993). 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Ageing Res Rev , 92, 102121. doi:10.1016/j.arr.2023.102121 Zhao, D., Liu, J., Wang, M., Zhang, X., & Zhou, M. (2019). Epidemiology of cardiovascular disease in China: current features and implications. Nat Rev Cardiol , 16(4), 203-212. doi:10.1038/s41569-018-0119-4 Supplementary Files GraphicalAbstract.jpg Cite Share Download PDF Status: Published Journal Publication published 06 Mar, 2025 Read the published version in Biotechnology Letters → Version 1 posted Editorial decision: Major revisions 02 Dec, 2024 Reviewers agreed at journal 31 Jul, 2024 Reviewers invited by journal 25 Jul, 2024 Editor assigned by journal 09 Jun, 2024 First submitted to journal 07 Jun, 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. 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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-4549466","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":331838546,"identity":"df789fc7-49cd-4de3-bfb9-fb7a29c5a33a","order_by":0,"name":"Yanyan Liu","email":"","orcid":"","institution":"Changchun University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Yanyan","middleName":"","lastName":"Liu","suffix":""},{"id":331838547,"identity":"671af499-031a-4326-9347-32f7d318756c","order_by":1,"name":"Meijiao Li","email":"","orcid":"","institution":"Changchun University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Meijiao","middleName":"","lastName":"Li","suffix":""},{"id":331838548,"identity":"36032727-8b18-45f6-90db-8c79306c9b88","order_by":2,"name":"Hao Zhang","email":"","orcid":"","institution":"Changchun University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Zhang","suffix":""},{"id":331838549,"identity":"73e81222-8416-47a7-9410-b8f871476d17","order_by":3,"name":"Le Gao","email":"","orcid":"","institution":"Changchun University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Le","middleName":"","lastName":"Gao","suffix":""},{"id":331838550,"identity":"3f86df0f-207f-4bd1-8a65-34071f462b9f","order_by":4,"name":"Jitao Liu","email":"","orcid":"","institution":"Jilin Getein Biotechnology Co., Ltd","correspondingAuthor":false,"prefix":"","firstName":"Jitao","middleName":"","lastName":"Liu","suffix":""},{"id":331838551,"identity":"0d1628eb-835f-438a-82ef-7b06218ef471","order_by":5,"name":"Yue Hou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxklEQVRIiWNgGAWjYBACAwhlA+WyEa8lDYiZSdNymAQt5uw9ZtI8FeflDc6fP8DwoewwA//sBvxaLHvOALWcuW244UYyA+OMc4cZJO4cIOCwGzlm0rxttxMMbjAzMPO2HWYwkEggRsu/cwkG5w8zMP8lXkvDgQSDA8kMzIxEaTlzrNhyzrFkw5k3kg0O9pxL55G4QUjL8eaNN97U2MnznT/48MGPMms5/hkEtAABiwSMdQCIeQiqBwLmD8SoGgWjYBSMghEMABOLQSsz0zyeAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0009-0000-5054-4493","institution":"Changchun University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Yue","middleName":"","lastName":"Hou","suffix":""},{"id":331838552,"identity":"29fa7954-72a0-4d8d-8a5f-04b4c84bc369","order_by":6,"name":"Jiancheng Xu","email":"","orcid":"","institution":"The First Hospital of Jilin University","correspondingAuthor":false,"prefix":"","firstName":"Jiancheng","middleName":"","lastName":"Xu","suffix":""}],"badges":[],"createdAt":"2024-06-08 07:47:41","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4549466/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4549466/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10529-025-03564-w","type":"published","date":"2025-03-06T15:57:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":63021160,"identity":"309c0c08-746b-45c0-9d40-c665cac31074","added_by":"auto","created_at":"2024-08-22 07:39:00","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":108487,"visible":true,"origin":"","legend":"\u003cp\u003eLp (a) calibration curve of immunolatex turbidity method. The absorbance displays the average value of 5 calibration samples±2 SD and the concentration of the calibration samples.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4549466/v1/6b79fdbaa76f32971b3451bb.jpg"},{"id":63021159,"identity":"6a6b1a52-a518-4137-8fad-ffcedd55538e","added_by":"auto","created_at":"2024-08-22 07:39:00","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":145168,"visible":true,"origin":"","legend":"\u003cp\u003eComparison results of serum samples between Lp (a) immunolatex reagent and maccure reagent\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4549466/v1/61ebf6a71136bba048818ea4.jpg"},{"id":78190473,"identity":"42127cac-7bc8-4a2c-9889-e6336b98aeb1","added_by":"auto","created_at":"2025-03-10 19:49:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":989194,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4549466/v1/f8fd39bb-c3b5-4e6d-83d4-8a8ff83370f0.pdf"},{"id":63021158,"identity":"add8d7aa-84d2-44ae-988d-de6c1f2f6294","added_by":"auto","created_at":"2024-08-22 07:39:00","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":110302,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4549466/v1/cbebb2308a8427caec30a5db.jpg"}],"financialInterests":"","formattedTitle":"Development of serum lipoprotein (a) detection using latex enhanced immunoturbidimetry","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAtherosclerosis is the root cause of most cardiovascular diseases (CVD), which causes more than 17.9\u0026nbsp;million deaths every year (Murray, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In recent years, although significant progress has been made in the prevention and treatment of CVD, it is still the main cause of incidence rate and mortality (Bułdak, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Mensah, Roth, \u0026amp; Fuster, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Timmis et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Xu, Murphy, Kochanek, \u0026amp; Arias, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhao, Liu, Wang, Zhang, \u0026amp; Zhou, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The occurrence of CVD is related to many factors, such as obesity, type 2 diabetes (T2DM) and metabolic syndrome (Mensah et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In addition, old age, hypertension, dyslipidemia, smoking and lack of exercise are traditional risk factors leading to accelerated atherosclerosis and early cardiovascular events (Yang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGenetic susceptibility is a key factor in the risk and progression of CVD. Although many standardized management methods have been used to address CVD risk factors, there are still significant CVD risks in some populations. Elevated levels of lipoprotein (a) (Lp (a)) in plasma are an independent and primarily gene determined pathogenic risk factor for CVD (Duarte Lau \u0026amp; Giugliano, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Ward et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In 1963, K \u0026aacute; re Berg first demonstrated that Lp (a) is a unique lipoprotein similar to low-density lipoprotein (LDL) (Berg, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1963\u003c/span\u003e). Lp (a) can accumulate in the subendothelial space and has the potential to cause thrombosis and atherosclerosis (Rath et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). Recent experiments have shown that high levels of Lp (a) lead to a higher incidence of CVD, accelerate disease progression, and increase the mortality rate of CV (Kamstrup, Benn, Tybjaerg-Hansen, \u0026amp; Nordestgaard, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Lampsas et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Ruscica, Sirtori, Corsini, Watts, \u0026amp; Sahebkar, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In addition, due to the lack of standardized Lp (a) measurement methods, there is diagnostic uncertainty in this field (Lampsas et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLatex enhanced turbometric immunoassay (LETIA) is an immunoassay technique that uses microparticles as markers. The core principle of this method is based on antigen antibody reactions, using latex coated with antibodies to react with specific analytes. Trigger the relationship between particle aggregation and analyte concentration through fast and simple measurement methods such as turbidimetry (Duan et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Xia et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In this method, the carrier particles used are polystyrene latex microspheres of varying diameters (Xia et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), which can bind to protein molecules through appropriate coupling methods. The content of the substance to be detected in the sample can be determined by detecting the degree of reduction in the transmittance of the solution through a spectral analyzer at a specific wavelength (Conde-S\u0026aacute;nchez, Rold\u0026aacute;n-Fontana, Chueca-Porcuna, Pardo, \u0026amp; Porras-Gracia, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Dong et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Duan et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Favresse et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Lapić et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Latex enhanced immunoturbidimetric analysis has highly automated characteristics and is suitable for large-scale clinical sample detection.\u003c/p\u003e \u003cp\u003eThe objective of this study is to establish an Lp (a) utilizing latex-enhanced immunoturbidimetric analysis, adhering to guidelines set forth by the Clinical and Laboratory Standards Institute (CLSI) (Plebani \u0026amp; Lippi, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This reagent kit offers the benefits of automated analysis, precision, rapidity, and resilience to interference. Below, we outline the characteristics and performance of this method, essential for validating its efficacy and establishing its reliability in clinical applications.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eInstrument\u003c/h2\u003e \u003cp\u003eAll measurements were performed with Hitachi 7100 fully automated biochemical analyzer (HITACHI, Japan).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSamples\u003c/h2\u003e \u003cp\u003eThe samples used in this study were artificial serum matrix (Braveds Biotechnology Co., Ltd., Shenzhen, China) and biochemical quality control samples (2833CH, 2835CH, Randox Company, UK) with different concentrations of recombinant antigens added (China, Yuansheng Biotechnology (Shanghai) Co., Ltd.) for this study, and there is no risk of clinical sample use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eLp (a) antibody\u003c/h2\u003e \u003cp\u003eThe rabbit polyclonal antibody targeting human Lp (a) is produced by Jianzhong Biotechnology Co., Ltd. (Ningbo, China). The antibody exhibited no cross-reactivity with apolipoprotein B and fibrinogen and was stored at 4℃.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eCalibration products\u003c/h2\u003e \u003cp\u003eLp (a) antigen (China, Yuansheng Biotechnology (Shanghai) Co., Ltd., concentration 2000mg/L) was used as a calibration sample, diluted with purified water to prepare standard solutions with concentrations of 0, 300, 600, 900, 1200, and 1500mg/L. In addition, Randox Lp (a) quality control products (batch numbers 2833CH, 2835CH, UK, Randox company) were used as secondary standards. The concentration of Lp (a) (295mg/L, 446mg/L) was repeated using latex enhanced immunoturbidimetric analysis (5 days, n\u0026thinsp;=\u0026thinsp;10 per day). The calibration sample was stored in a plastic bottle at -30℃ or freeze-dried under 600 mmol/L sucrose conditions (Borque, Maside, Rus, \u0026amp; del Cura, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eLatex reagents\u003c/h2\u003e \u003cp\u003e10% of solid carboxylated polystyrene latex particles with a diameter of 95nm (JSR-P0011), purchased from JSR Corporation in Japan, were used. Following the method described in the literature, covalent coupling was performed between the Lp (a) antibody and JSR-P0011 latex particles (Borque et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). The obtained 0.5% solid coated particle solution was stored at 4℃. The original solution should be frozen or freeze-dried for long-term storage.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMeasurement procedure\u003c/h2\u003e \u003cp\u003eOn the Hitachi 7100 fully automatic biochemical analyzer, the quantitative addition of samples and reagents was meticulously carried out. A comprehensive 6-point calibration curve was meticulously constructed, spanning concentrations from 0 to 1500 mg/L, with each concentration meticulously tested thrice for precision. The procedural steps are outlined as follows:\u003c/p\u003e \u003cp\u003eCommence by dispensing 200 \u0026micro;L of glycine buffer (comprising 0.1 mol/L glycine, 0.15 mol/L NaCl, 10 g/L bovine serum albumin, 30 g/L polyethylene glycol, and 0.6 mmol/L sodium azide, pH adjusted to 8.2). Subsequently, introduce 50 \u0026micro;L of latex reagent into the buffer to initiate the reaction. Follow by adding 3 \u0026micro;L of the experimental sample to the reaction mixture. Proceed to monitor the ensuing reaction at a wavelength of 600 nm over a duration of 5 minutes, meticulously maintaining the temperature at 37\u0026deg;C.\u003c/p\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003eComparison with commercially available reagent kits\u003c/h2\u003e \u003cp\u003eCompare the performance of the Lp (a) immunolatex reagent with similar products available in the market (Maccure, China, Sichuan Mike Biotechnology Co., Ltd.) by conducting linear fitting and scatter plot analyses on the test results obtained from 50 serum samples within the testing range.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003eInterference substances\u003c/h2\u003e \u003cp\u003eThe interference study followed the Glick program (Glick, Ryder, \u0026amp; Jackson, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1986\u003c/span\u003e), adding different concentrations of antigens to the artificial serum matrix to prepare basic samples. By adding ascorbic acid (200, 400, 800, 1000, 2000 mg/L, China, Ningbo National Pharmaceutical Group Chemical Reagent Co., Ltd.), fat emulsion (0.25, 0.50, 1.00, 1.50, 2.00%, China, Anhui Fengyuan Pharmaceutical Co., Ltd.), bilirubin (100, 200, 400, 600, 800 \u0026micro; Mol/L, China, Shanghai Alfa Aesar (China) Chemical Co., Ltd.) and hemoglobin (0.5, 1.0, 2.0, 4.0, 6.0g/L, China, Ningbo National Pharmaceutical Group Chemical Reagent Co., Ltd.) were added to the base sample. Each sample was tested three times, and the average value was taken to calculate the interference rate and evaluate its impact on the measurement results.\u003c/p\u003e \u003cp\u003eThe interference rate is calculated as the interference concentration divided by the base sample concentration, multiplied by 100%. An interference rate within the range of \u0026plusmn;\u0026thinsp;10% indicates no significant interference, while a rate exceeding\u0026thinsp;\u0026plusmn;\u0026thinsp;10% signifies significant interference.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eStatistics\u003c/h2\u003e \u003cp\u003eTo investigate the relationship between latex-enhanced immunoassay and commercially available test kits, we will undertake Passing-Bablok regression and Bland-Altman analysis (Fu et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These methods will help assess the agreement and bias between the two measurement techniques. Additionally, we will calculate the correlation coefficient of Lp (a) values using linear regression analysis. Furthermore, samples at three concentration levels will be utilized to evaluate deviation, coefficient of variation (CV), and recovery rate, providing insights into accuracy, precision, and recovery efficiency across different concentrations.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eCalibration curve\u003c/h2\u003e \u003cp\u003eLp (a) measurement was conducted using a 6-point calibration program on the Hitachi 7100 fully automated biochemical analyzer. The analysis range extends from 0 to 1500 mg/L (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The linear equation demonstrates a strong linear relationship between each concentration and the detection results, encompassing both normal and pathological serum values. In case the concentration exceeds 1500 mg/L, physiological saline was diluted fourfold and subsequently analyzed automatically by the instrument. The stability of the calibration curve is guaranteed for up to 30 days when reagents are stored at 2 to 8\u0026deg;C.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eInaccuracy\u003c/h2\u003e \u003cp\u003eConduct 20 consecutive measurements of the Randox quality control substance (2833CH) and observe the imprecision density (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The coefficient of variation within the batch is 1.10%, indicating excellent repeatability in measurement results. Furthermore, when measuring the same Randox quality control sample (2835CH) over 20 consecutive days, the inter-batch coefficient of variation was 1.79%, demonstrating the immunolatex turbidity method's accuracy (Fu et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2024\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\u003eResults of imprecision measurement for Lp (a) immune latex\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\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=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProject Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRandox Quality Control (2833CH)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRandox Quality Control (2835CH)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMeasurement frequency\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAverage value (\u0026oline;χ)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e304.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e464.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaximum value (\u003cem\u003eX\u003c/em\u003e \u003csub\u003emax\u003c/sub\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e306.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e476.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMinimum value (\u003cem\u003eX\u003c/em\u003e \u003csub\u003emin\u003c/sub\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e303.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e453.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStandard deviation (SD) (\u003cem\u003eSD\u003c/em\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCoefficient of variation (\u003cem\u003eCV\u003c/em\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.10%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.79%\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 \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eSensitivity\u003c/h2\u003e \u003cp\u003eTo assess the linear relationship, continuously dilute two distinct Randox quality control samples (2833CH, 2835CH). The linear regression coefficient (R\u003csup\u003e2\u003c/sup\u003e) between Lp (a) and dilution for all samples is 0.994. Within the measurement range, the deviation between the measured value and the theoretical value must not exceed 5%. To determine the lower detection limit, calculate the average value of 20 repeated measurements of physiological saline solution (zero signal) and add 3 standard deviations, resulting in a lower detection limit of 7mg/L.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eInterference substances\u003c/h2\u003e \u003cp\u003eInterference samples of varying concentrations are measured with the sample concentration serving as the reference value. When the sample's ascorbic acid concentration is \u0026le;\u0026thinsp;2000 mg/L, fat emulsion concentration is \u0026le;\u0026thinsp;2.00%, bilirubin concentration is \u0026le;\u0026thinsp;800 \u0026micro;mol/L, and hemoglobin concentration are \u0026le;\u0026thinsp;6g/L, the interference rate of interferents on Lp (a) determination should be \u0026le;\u0026thinsp;\u0026plusmn;\u0026thinsp;10% (refer to Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This demonstrates the absence of endogenous interference in the determination of the immunolatex reagent.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDistribution of interference rates (%) of four substances\u003csup\u003e★\u003c/sup\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\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=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eAscorbic acid\u003c/p\u003e \u003cp\u003e(mg/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eFat emulsion\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eBilirubin\u003c/p\u003e \u003cp\u003e(\u0026micro;mol/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eHemoglobin\u003c/p\u003e \u003cp\u003e(g/L)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u0026thinsp;2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-2.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u0026thinsp;0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u0026thinsp;4.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-4.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u0026thinsp;2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-3.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u0026thinsp;5.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e4.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-7.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-8.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-5.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u0026thinsp;4.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e6.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-9.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"11\"\u003e\u003csup\u003e★\u003c/sup\u003e A: Concentration; B: Interferencerate\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eRecovery rate of spiking\u003c/h2\u003e \u003cp\u003eRandox quality control (2835CH) with known concentration was added to 5 mixed serum samples and measured. The results of the spiked recovery rate are shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, with a range of 96.5% -105.9%. The average recovery rate is 100.98%.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRecovery rate of Lp (a) immunolatex determination\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=\"char\" char=\".\" 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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHeoretical spiked value\u003c/p\u003e \u003cp\u003e(mg/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMeasured value\u003c/p\u003e \u003cp\u003e(mg/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRecovery rate\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e129.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e125.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e97.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e171.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e182.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e105.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e361.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e354.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e97.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e439.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e453.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e103.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e538.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e519.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e96.5\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 \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eCorrelation coefficient\u003c/h2\u003e \u003cp\u003eUsing the Hitachi 7100 automatic biochemical analyzer, 50 serum samples were simultaneously tested with the Lp (a) immunolatex assay reagent and the control Lp (a) immunolatex reagent (from Maccure). The method comparison results are displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe serum Lp (a) values ranged from 9.1 to 1041.1 mg/L. There was a strong correlation observed between the immune latex reagent and the Maccure reagent samples, with no significant difference noted (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eResearch shows that Lp (a) is an independent risk factor for atherosclerosis and thrombosis (Bułdak, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Mensah et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Timmis et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Yang et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). However, currently there are methodological differences in Lp (a) detection in clinical laboratories due to issues with methods and standardization. Traditional Lp (a) measurement methods typically involve cumbersome manual analysis techniques. These methods generally suffer from issues such as unstable precision and long detection time. In order to overcome these problems, several automatic immune turbidity analysis methods have been developed in recent years (Borque, Rus, del Cura, Maside, \u0026amp; Escanero, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Tiran, Tiran, Hojas, Kostner, \u0026amp; Wilders-Truschnig, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), but there are significant differences in the detection results between these methods (Borque, Maside, \u0026amp; Iglesias, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). One of the main issues in detecting serum Lp (a) in the past was the difference in the purity of Lp (a) antibodies, as well as the difference in particle size selection due to carboxylated polystyrene microspheres (Tiran et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis study utilizes carboxylated latex particles, 95 nm in diameter, to covalently coat Lp (a) antibodies. The sample and coated latex particles underwent a 5-minute incubation at 37\u0026deg;C, followed by measuring turbidity change at 600 nm to quantify the agglutination reaction. Optimal detection efficacy is achieved with a coupling ratio of 5:1 between latex microspheres and Lp (a) antibody, resulting in larger signal generation due to the smaller volume of microspheres.\u003c/p\u003e \u003cp\u003eThis analysis method offers rapid, accurate, and fully automated measurements on the Hitachi 7100 automatic biochemical analyzer. Using Randox biochemical quality control samples, precision evaluation revealed an intra-batch precision CV% of 1.10% and an inter-batch precision CV% of 1.79%. The detection limit (LoD) for Lp (a) is 7 mg/L, with a correlation coefficient R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9946 within the 0-1500 mg/L concentration range. Notably, interference from bilirubin, fat emulsion, hemoglobin, and ascorbic acid is absent. When compared with a commercially available latex-enhanced particle immunoturbidimetric Lp (a) assay reagent, the developed reagent exhibited a strong correlation (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9972) on the Hitachi 7100 analyzer. Overall, this method demonstrates superior comprehensive performance compared to previously reported Lp (a) measurement methods.\u003c/p\u003e \u003cp\u003eIn conclusion, our study confirms the viability of latex-enhanced particle immunoturbidimetry for serum Lp (a) determination on the Hitachi 7100 fully automated biochemical analyzer. This method demonstrates speed, excellent linearity, precision, and full automation. Through rigorous performance verifications, the particle enhancement method meets expected criteria, thus proving its potential for widespread application in clinical and related fields for Lp (a) detection.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eConceptualization: YY L, Y H and JC X; Data curation: YY L, MJ L, Y H and JT L; Formal analysis: YY L, L G, Y H and JC X; Funding acquisition: H Z, Y H and JC X; Methodology:YY L, H Z, L G and Y H; Visualization: YY L, MJ L and JT L; Writing original draft: YY L, MJ L and JT L; Writing - review \u0026amp;editing: YY L, H Z, L G ,JC X and Y H; Supervision: Y H and JC X.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u0026nbsp;\u003c/strong\u003eThe authors have no potential conflicts of interest. Special thanks to Dr. Lipeng Xu from Jilin Getein Biotechnology Co., Ltd. for his valuable assistance during the experiment. This work was supported by Science and Technology Department of Jilin Province (Grant No.20220401085YY).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConfict of interest\u0026nbsp;\u003c/strong\u003eThe authors have no relevant fnancial or non-fnancial interests to disclose.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e This article does not contain any studies with human participants or animals performed by any of the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e This article does not contain any studies with human participants or animals performed by any of the authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to publish\u003c/strong\u003e This article does not contain any studies with human participants or animals performed by any of the authors\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003eThe data generated in this study are available upon request from the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e The authors declare that there are no confict of interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBerg, K. 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Scientific reports\u003cem\u003e, \u003c/em\u003e7, 40090. doi:10.1038/srep40090\u003c/li\u003e\n\u003cli\u003eXu, J., Murphy, S. L., Kochanek, K. D., \u0026amp; Arias, E. (2022). Mortality in the United States, 2021. NCHS Data Brief(456), 1-8.\u003c/li\u003e\n\u003cli\u003eYang, K., Hou, R., Zhao, J., Wang, X., Wei, J., Pan, X., \u0026amp; Zhu, X. (2023). Lifestyle effects on aging and CVD: A spotlight on the nutrient-sensing network. Ageing Res Rev\u003cem\u003e, \u003c/em\u003e92, 102121. doi:10.1016/j.arr.2023.102121\u003c/li\u003e\n\u003cli\u003eZhao, D., Liu, J., Wang, M., Zhang, X., \u0026amp; Zhou, M. (2019). Epidemiology of cardiovascular disease in China: current features and implications. Nat Rev Cardiol\u003cem\u003e, \u003c/em\u003e16(4), 203-212. doi:10.1038/s41569-018-0119-4\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":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"biotechnology-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bile","sideBox":"Learn more about [Biotechnology Letters](https://www.springer.com/journal/10529)","snPcode":"10529","submissionUrl":"https://submission.nature.com/new-submission/10529/3","title":"Biotechnology Letters","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Cardiovascular disease, Lipoprotein (a), Measurement value, Atherosclerosis, Latex enhanced turbidimetric immunoassay, Abnormal blood lipids","lastPublishedDoi":"10.21203/rs.3.rs-4549466/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4549466/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eLipoprotein (a) (Lp (a)) is indeed a significant factor in cardiovascular health, as it is a product of low-density lipoprotein cholesterol-like particles that bind to apolipoprotein (a). Elevated levels of Lp (a) have been linked to an increased risk of cardiovascular diseases (CVD), hastening disease progression and raising CVD mortality rates. However, the absence of standardized measurement methods for Lp (a) contributes to diagnostic uncertainties.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003eA quantitative measurement method for serum Lp (a) was developed using fully automated latex-enhanced particle immunoturbidimetry technology represents a significant advancement in diagnostic capabilities. The key parameters such as repeatability, stability, linearity, and method comparison were evaluated to ensure the accuracy of the assay.\u003c/p\u003e\u003ch2\u003eResult\u003c/h2\u003e \u003cp\u003eThe Lp (a) in samples was recognized by carboxylated latex particles covalently coated with anti-Lp (a) antibodies. The content of Lp (a) was quantified by measuring the changes in turbidity generated by agglutination at 600 nm. With precision CV% within the batch of 1.10% and inter-batch precision CV% of 1.79%, it demonstrates reliable performance using Randox biochemical quality control samples. The detection limit of 7 mg/L and a high correlation coefficient (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9946) at concentrations of 0-1500 mg/L further validate its effectiveness.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe quantitative determination method of serum Lp (a) based on latex-enhanced immunoturbidimetric analysis indeed provides rapid results, high accuracy, and automation, making it suitable for routine clinical testing. This method relies on the interaction between Lp (a) and latex particles, allowing for efficient measurement in serum samples.\u003c/p\u003e","manuscriptTitle":"Development of serum lipoprotein (a) detection using latex enhanced immunoturbidimetry","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-22 07:38:55","doi":"10.21203/rs.3.rs-4549466/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revisions","date":"2024-12-02T13:16:05+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-07-31T13:27:03+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-07-25T17:22:21+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-06-09T06:49:22+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biotechnology Letters","date":"2024-06-08T03:47:25+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biotechnology-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bile","sideBox":"Learn more about [Biotechnology Letters](https://www.springer.com/journal/10529)","snPcode":"10529","submissionUrl":"https://submission.nature.com/new-submission/10529/3","title":"Biotechnology Letters","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"266d662a-e5e8-4a6b-883c-2e3c978e855e","owner":[],"postedDate":"August 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-03-10T19:45:20+00:00","versionOfRecord":{"articleIdentity":"rs-4549466","link":"https://doi.org/10.1007/s10529-025-03564-w","journal":{"identity":"biotechnology-letters","isVorOnly":false,"title":"Biotechnology Letters"},"publishedOn":"2025-03-06 15:57:56","publishedOnDateReadable":"March 6th, 2025"},"versionCreatedAt":"2024-08-22 07:38:55","video":"","vorDoi":"10.1007/s10529-025-03564-w","vorDoiUrl":"https://doi.org/10.1007/s10529-025-03564-w","workflowStages":[]},"version":"v1","identity":"rs-4549466","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4549466","identity":"rs-4549466","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}