Development and Clinical Application of Component-Resolved Diagnostic Using Light-Initiated Chemiluminescence Assay to characterize House Dust Mite Components-Specific IgE

Development and Clinical Application of Component-Resolved Diagnostic Using Light-Initiated Chemiluminescence Assay to characterize House Dust Mite Components-Specific IgE | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 30 June 2025 V1 Latest version Share on Development and Clinical Application of Component-Resolved Diagnostic Using Light-Initiated Chemiluminescence Assay to characterize House Dust Mite Components-Specific IgE Authors : Qinqin Liu , Yuanmin Sun , Huiqiang Li , and Xue-Yan Wang 0000-0003-3908-3862 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.175131003.34249593/v1 Published Journal of Immunological Methods Version of record Peer review timeline 143 views 90 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Abstract Objective: Developed a rapid, sensitive, and homogeneous immunoassay to characterize specific immunoglobulin E (sIgE) against HDM allergens in serum samples. Methods: Initially, the reaction conditions were optimized, and the levels of sIgE specific to the major HDM components-Der p 1, Der p 2, and Der p 23-were measured using the Light-initiated Chemiluminescence Assay (LiCA) assay. The performance of this assay was evaluated in accordance with established clinical guidelines. Subsequently, we analyzed the distribution of these components among 90 children with various allergic diseases. Results: We established an optimal incubation time of 45 minutes. The coefficient of variation (CV) for repeatability ranged from 2.64% to 9.36%, while the intermediate precision varied from 5.77% to 9.89%. Component-resolved diagnosis (CRD) indicated that Der p 2 was the most frequently recognized componen (75.56%), followed by Der p 1 at 64.44%. The highest co-sensitization rate was observed between Der p 1 and Der p 2 (35.56%). Der p 23 sIgE levels were significantly elevated in patients with AA (P < 0.001). Additionally, a greater complexity in allergic symptoms was associated with an increased positive rate of Der p 23 (P = 0.007). Conclusion:We established an ssay for HDM component-sIgE using LiCA technology. Der p 2 was identified as the most frequently recognized allergen among the HDM components. Furthermore, our findings indicate a significant correlation between the complexity of allergic symptoms and elevated levels of Der p 23 sIgE. Introduction Allergic diseases, characterized by an abnormal immune response to allergens, encompass conditions such as AR, AA, and AD 1 . The global prevalence of allergic diseases is estimated to range from 10% to 40% 2-4 . In developed countries, asthma affects approximately 8% of the population 5 , while AR impacts about 40% globally 6 . Among children, the lifetime prevalence of doctor-diagnosed AD is reported to range from 4.7% to 20.2% when assessed at ages 7 to 15 years 7 . Over the past three decades, the prevalence of AD among Chinese children aged 1 to 7 has reached 12.94% 8 . Furthermore, there has been a notable increase in the co-occurrence of multiple allergic conditions within the same patient, which complicates disease management 9 . Early diagnosis of patients with allergies is essential for avoiding allergens and guiding clinical treatment. Currently, the diagnosis of allergic diseases relies on patients’ clinical history, physical examination, and allergen testing, with the latter being crucial for accurate diagnosis 10 . The commercially available sIgE kits commonly used in practice are derived from natural allergen extracts, which comprise a heterogeneous mixture of not only major allergens but also cross-reactive proteins, non-allergenic antigens, and interfering substances 9 . These factors can compromise the sensitivity and specificity of the tests and contribute to cross-reactivity issues in the diagnosis of polysensitized patients 11 . In recent years, the rapid advancement of clinical and basic research in the field of molecular allergy has facilitated the gradual application of allergen CRD 11-13 . Molecular diagnosis and monitoring of allergen-specific antibody responses may enhance the efficacy of specific immunotherapy in real world settings through a personalized approach 14 . HDM are prevalent in our living environment and represent one of the most prevalent inhalant allergens 15 . Reports indicate that over 50% of individuals with allergies are sensitized to HDM, and up to 85% of children diagnosed with AA exhibit sensitization to HDM 16 . More than 40 distinct components of HDM have been identified. Considering the significant differences in molecular sensitivities to HDM across various regions and populations, CRD for HDM is essential for accurately identifying allergenic components. This approach aids in understanding the importance of allergen avoidance, the application of HDM-specific immunotherapies, and provides deeper insights into clinical symptoms, thereby facilitating the diagnosis and management of allergic diseases 14,17 . Cohort studies have indicated that Der p 1, Der p 2, and Der p 23 are the primary components of HDM allergens, with a prevalence of allergy exceeding 50% 18 . However, the current in vitro diagnostic techniques for serum-sIgE antibodies to HDM allergens in China predominantly utilize crude extracts, such as enzyme-linked immunosorbent assay (ELISA), fluorescent enzyme immunoassay (FEIA), and Dot-ELISA. These methods lack the specificity necessary to accurately identify individual HDM allergenic components 19 . Accurate diagnosis using purified or recombinant allergen components is crucial for individuals suffering from HDM allergies. Various methods are available for the detection of HDM component allergens. For example, the ImmunoCAP ISAC 112 system can simultaneously detect 112 different allergenic components 20 ; however, it is not yet accessible in China. Additionally, detection systems developed by Changsha Haike Biotechnology Co., Ltd. and Hangzhou Zhejiang Dixon Bio-genetic Engineering Co., Ltd. have encountered methodological limitations that have hindered their widespread adoption in clinical practice. Consequently, this study establishes a LiCA-based method that quantitatively detects sIgE antibodies to multiple HDM allergen components. The performance of this method was validated according to clinical guidelines to ensure compliance with the requirements of clinical laboratories. Finally, we analyzed the distribution of these components in children with various allergic diseases. Establishment of the LiCA Method for HDM Component-sIgE Reagents and Chemicals Three extracts of HDM allergen components were obtained from Beijing BioNC Biotech Co. (Beijing, China). Biotinylated mouse anti-human IgE antibodies were sourced from Abcam (Cambridge, UK). Bare chemibeads and streptavidin-coated sensibeads were supplied by Beyond Biotech (Shanghai, China). Coupling of Allergens with Beads Coupling of Aldehyde Microspheres The HDM component antigens were dissolved in a 0.1 M carbonate buffer solution at pH 9.5. Prior to use, 1 mg of chemibeads was washed twice with purified water, then resuspended in 0.1 M CBS (pH 9.6), vortexed for 1 minute, and sonicated for 30 seconds. The antigen solutions were subsequently added and incubated on a rotary shaker at 37°C overnight. The following day, an 8 mg/mL sodium borohydride solution was introduced to the mixture and shaken for 2 hours at room temperature. Cross-linking was terminated by adding 75 mg/mL glycine, followed by an additional hour of shaking at room temperature. Finally, the HDM component antigen-coated aldehyde chemibeads were washed, and the remaining pellets were resuspended in 200 μL of storage buffer (20 mM HEPES + 0.2% BSA + 0.1% Proclin-300) for storage. Coupling of Carboxy Microspheres HDM component antigens were dialyzed overnight against a 0.02 M HEPES buffer. The chemibeads were resuspended in a 0.05 M MES buffer (pH 6.0), and the supernatant was discarded following centrifugation at 10,000×g for 2 minutes. The chemibeads were then resuspended in 80 mL of activation buffer, combined with 0.5 mg of sulfonyl chloride solution and 0.5 mg of EDC solution, and stirred at room temperature for 20 minutes to activate the carboxyl groups on their surface. After washing with PBS, the component antigens were added, and incubation was carried out with oscillations at 37℃ for 4 hours. The beads were subsequently resuspended in a blocking buffer containing 15% BSA and incubated on a rotary shaker at room temperature for 14 hours to block any remaining active sites. Following washing, the HDM component antigen-coated carboxylated chemibeads were resuspended at a concentration of 10 mg/mL and stored at 4℃. Selection of Allergen-coupled Chemibeads The selection of chemibeads significantly impacted the antigenic activity on the microsphere surface after coupling with the three HDM components. To evaluate this, three serum pools-comprising high, medium, and low levels of Der p sIgE antibodies-were chosen, with serum samples from non-HDM-allergic individuals serving as negative controls. Each serum pool was evaluated using three types of antigen-coated carboxyl and aldehyde-based chemibeads to detect chemiluminescence (CL) signals and calculate signal-to-noise ratios. From these results, the optimal chemibeads for each antigen were identified. The coupling of HDM components with chemibeads was conducted following a previously established standard procedure 21 . Optimization of Reaction System The incubation time during the initial reaction phase and the buffer system were the primary factors influencing the performance of LiCA. Initially, six concentrations were prepared using high and low positive serum samples containing Der p 1, Der p 2, and Der p 23-sIgE antibodies, which were mixed in varying ratios. The first-stage incubation times were established at 30 minutes, 45 minutes, and 60 minutes, respectively. Calibration Curve The World Health Organization’s 3rd Generation Human sIgE International Reference Product (11/234) served as the reference material for the HDM-sIgE calibrator. This calibrator was diluted to concentrations of 100 kUA/L, 50 kUA/L, 17.5 kUA/L, 3.5 kUA/L, 0.75 kUA/L, and 0.35 kUA/L. Serum from non-allergic individuals was utilized as the 0 kUA/L control. All seven calibration points were analyzed using LiCA assays. A calibration curve was generated based on four-parameter logistic regression, employing the concentrations and corresponding signal values, with results quantified according to the various HDM components tested. Precision According to the Clinical and Laboratory Standards Institute (CLSI) EP5-A2 guidelines, precision was evaluated through both repeatability and intermediate precision 22 . Three serum pools with varying levels of HDM-sIgE (high, medium, and low) were selected for this assessment. To evaluate repeatability, each specimen was tested 20 times within a single analytical run. For intermediate precision, the same specimens were measured 8 times daily over a span of 5 consecutive days. Patients From August 2023 to March 2024, this study collected data from children aged 1 to 18 years who were clinically suspected of having allergic diseases, as assessed by pediatricians at the Second Hospital of Tianjin Medical University. A total of 90 newly diagnosed patients with complete datasets were included for further analysis. Serum specimens with HDM-sIgE levels ≥0.35 kUA/L were considered positive, as measured by the ImmunoCAP system. Positive sIgE tests were categorized into six classes: Class 1 (≥0.35 to <0.70 kUA/L), Class 2 (≥0.70 to <3.50 kUA/L), Class 3 (≥3.50 to <17.50 kUA/L), Class 4 (≥17.50 to Class 6 (≥100.00 kUA/L) 23 . The study received approval from the Ethics Committee of the Second Hospital of Tianjin Medical University (TMUHMEC 2024K090) and was conducted in accordance with the principles outlined in the Declaration of Helsinki. The relationship between HDM components-sIgE antibody and clinical characteristics We examined the distribution characteristics and co-sensitization patterns of the three HDM components. Statistical analyses were conducted to evaluate the expression characteristics of these components in relation to patients’ demographic, clinical, and serological features. Importantly, we also analyzed the distribution of these components across various allergic diseases. Statistical analysis All statistical analyses were conducted using SPSS software, Version 25.0. Parametric quantitative data are expressed as the mean ± standard deviation (SD), while non-parametric quantitative data are presented as medians with interquartile ranges (IQR). Categorical data are reported as percentages, reflecting the proportion of positive results. Comparisons of proportions between groups were performed using chi-square tests (χ²) or Fisher’s exact test, as appropriate. F-tests were employed to assess variance differences among groups. Correlation analyses for non-parametric data were conducted using Spearman’s rank correlation tests, with correlation coefficients denoted as rs. A P-value of < 0.05 was considered statistically significant. Logistic regression curves and other graphical representations were generated using GraphPad Prism software, Version 9.5. Results Selection of Components for Bead Coupling Mode For the three HDM component antigens, the various chemibeads were the primary factor influencing the antigenic activity on the microsphere surface following coupling. Initially, three serum pools with high, medium, and low levels of Der p sIgE antibodies were selected. Subsequently, each serum pool was analyzed using the three components of antigen-coated carboxy and aldehyde-based chemical beads to assess the CL signals and calculate the signal-to-noise (S/N) ratios. Ultimately, Der p 1-carboxy microspheres, Der p 2-carboxy microspheres, and Der p 23-aldehyde microspheres were chosen for coupling and binding in preparation for subsequent detection (Figure 1). Optimization of Incubation Time The duration of the initial incubation during the reaction and the buffer system employed were the primary factors influencing the performance of LiCA. Initially, six concentrations were prepared using high and low positive serum samples containing Der p 1 and Der p 2-sIgE antibodies, which were mixed in various ratios. The incubation times for the gradients in the first stage of the study were set at 30 minutes, 45 minutes, and 60 minutes, respectively. As illustrated in Figure 2, the CL signal value exhibited a trend of initially increasing and then decreasing with the prolongation of the reaction time. Consequently, 45 minutes was determined to be the optimal reaction time for the first stage. The reaction time for the second stage was set at 10 minutes, resulting in a total reaction time of 55 minutes. Establishment of the LiCA Method for HDM Component-sIgE Construction of Calibration Curve The sIgE calibrator was diluted to concentrations of 100 kUA/L, 50 kUA/L, 17.5 kUA/L, 3.5 kUA/L, 0.75 kUA/L, and 0.35 kUA/L, while serum from non-allergic individuals was measured at 0 kUA/L. LiCA assays were conducted for all seven calibration points. A calibration curve was generated using 4-parameter logistic regression, incorporating the concentrations and corresponding chemical signal values. Performance Evaluation Precision To evaluate precision, three serum pools were analyzed using LiCA, and the mean, standard deviation (SD), and coefficient of variation (CV) were calculated. As presented in Table 1, the results of the repeatability study indicated that the CV ranged from 2.64% to 9.36%. In the intermediate precision study, the CV ranged from 5.77% to 9.89%. In both the repeatability and intermediate precision assessments, the CV values were below 15%, which is considered acceptable. Demographic Characteristics of Patients A total of 90 patients were screened in this study, comprising 66 males (73.33%) and 24 females (26.67%), with a mean age of 8.50±3.31 years (range: 3-17 years; interquartile range: 6.00-11.25 years). In our study, the positive values of the other two HDM components, sIgEs, in male patients were slightly higher than those in females, except for Der p 1; however, none of the disparities were statistically significant (P > 0.05). Children were categorized into four age groups based on developmental stages: 4 toddlers (>1-3 years), 33 preschool children (>3-7 years), 40 school-aged children (>7-12 years), and 13 adolescents (>12-18 years). We found that Der p 23-sIgE levels were significantly different among the four age groups, with the highest IgE level observed in the adolescent group (P = 0.003, χ² = 12.557). IgE levels of Der p 2 were also highest in the adolescent group. In contrast, the IgE level of Der p 1 was highest in the toddler group, although there were no significant differences among the four groups. The majority of the children had allergic rhinitis (94.44%), followed by allergic asthma (60%) and allergic skin disease (8.89%). Among the children, classes 3 and 4 were the most common, each accounting for 30%, followed by class 2 (27.78%). (Table 2). Expression Characteristics of HDM Component-sIgE Antibody Molecular Reactivity Patterns of HDM-allergic Patients Among the 90 Der p-positive children, 88 (97.78%) tested positive for at least one of the three major components. Specifically, 58 (64.44%) were sensitized to Der p 1, 68 (75.56%) to Der p 2, and 27 (30.00%) to Der p 23. The detection rate of Der p 2 in patients with a class 4 or higher sIgE response to Der p exceeded 90%. Additionally, a higher concentration of Der p-sIgE correlated with an increased rate of positivity for Der p 2 (P = 0.001, χ²= 18.371) (Table 2). As illustrated in Figure 3, we examined co-sensitization among the three HDM allergens. The results indicated that the highest co-sensitization rate within the HDM allergens group was between Der p 1 and Der p 2, with a total of 32 subjects exhibiting co-sensitization (35.56%). Furthermore, 13 (14.44%) of the patients were sensitized to all three components, while 39 (43.33%) were sensitized to two out of the three components. In our study population, 87.5% of patients with AD were sensitive to two or more components, in contrast to 57.5% in the AR group and 60.00% in the AA group. The mean number of identified components was 1.73±0.725 in AD patients, 1.64 ± 0.698 in AR patients, and 1.83 ±0.770 in AA patients, with AD patients exhibiting the highest average number of identified components at 2.00±0.535. However, there was no significant difference among the three groups (P = 0.155). Additionally, no significant difference was observed in the number of component sensitizations between patients with single allergic symptoms and those with multiple allergic symptoms (P = 0.295). Subsequently, we categorized patients into distinct groups based on the type and number of allergic diseases. The mean number of sensitized components was 2.33±0.866 in nine patients diagnosed with AA alone, 1.44±0.641 in 27 patients with AR alone, one in a single patient with AD alone, and 1.67±0.732 in 46 patients presenting with a combination of AA and AR. Additionally, the mean was 2.00 in the two patients who had both AR and AD concurrently. There were no cases of AA combined with AD; however, five patients exhibited a combination of AA with AR and AD, resulting in an overall average of component sensitization of 2.20 ± 0.447. We further investigated the patterns of IgE sensitization and the distribution of component IgE levels in relation to the presence of concomitant asthma. As illustrated in Figure 4, we found that Der p 23 IgE levels were significantly elevated in patients with asthma (45.00% vs. 0.00% in patients with and without asthma, respectively; P < 0.001). Similarly, the proportion of patients with IgE directed against Der p 23 was greater in those with concomitant AD (62.50% vs. 26.83% in patients with and without AD, respectively; P < 0.05). Based on the prevalence of allergic diseases, we categorized them into three groups: those with one disease, two diseases, and three diseases. A greater complexity in allergic symptoms was associated with an elevated positive rate of Der p 23 (P=0.007, χ 2 =9.117). Subsequently, we categorized patients into distinct groups based on the type and number of allergic diseases. The positive rate of Der p 23 was significantly higher in the group with AA+AR+AD compared to those with AA only, AR alone, AD alone, AA+AR, and AR+AD (P0.05). Discussions The characterization of HDM components is crucial for accurately identifying allergenic components, formulating HDM immunotherapy regimens, and enhancing the effectiveness of specific immunotherapy 17 . However, there are still limited methods available for detecting HDM sensitizing components. LiCA, a nanosphere-based homogeneous immunoassay, offers high sensitivity, reproducibility, and rapidity, making it particularly suitable for the detection of allergen-sIgE 24 . Published studies have emphasized the advantages of LiCA technology compared to current clinical methods for the quantitative detection of allergen sIgE 21, 24-27 . This approach addresses the limitations of enzyme stability in ELISA and the prolonged turnaround time associated with FEIA, while effectively measuring varying antibody concentrations with minimal sample volume. In this study, we developed an indirect LiCA method for detecting three components of HDM allergens: Der p 1, Der p 2, and Der p 23-sIgE. This method requires only 5.0μL of serum for each test, which is significantly lower than the 100μL required by the Immuno CAP system. Consequently, the LiCA method is particularly suitable for specimens obtained from infants and children. Furthermore, LiCA demonstrates excellent sensitivity and repeatability. Our study demonstrated that 90% of the subjects were sensitive to multiple allergen components. Der p 2 exhibited the highest sensitization rate at 75.56%, followed by Der p 1 at 64.44%. These results are consistent with the findings reported by Gan H et al 28 . 27 patients (30%) exhibited sensitivity to Der p 23, which is higher than the reported rate of Der p 23 positivity in Northern China (23.3%) as noted by Gan H et al 28 . A study indicated that Der p 23 was the most prominent monosensitizer among the subjects sensitized to HDM, with 8.2% of these individuals responding exclusively to Der p 23, which is consistent with our findings 29 . Previous research has demonstrated that age influences IgE levels 30 . Our results show that the IgE level for Der p 23 is highest in the adolescent group (P=0.003, χ 2 =12.557). Monosensitization to the HDM allergen components Der p 1, Der p 2, and Der p 23 was observed in 43.33% of HDM-allergic patients. Among these, 10 patients (11.11%) tested positive solely for Der p 1, 19 patients (21.11%) for Der p 2, and seven patients (7.78%) for Der p 23. We subsequently investigated co-sensitization among the three HDM allergens, which revealed that the highest co-sensitization rate occurred between Der p 1 and Der p 2, with a total of 32 subjects (35.56%). This finding may be attributed to the high homology between 31 . We found that Der p 23 IgE levels were significantly elevated in asthma patients (45.00% vs 0.00%, P<0.001). Similarly, the proportion of patients with IgE specific to Der p 23 was greater in those with concomitant AD (62.50% vs 26.83%, P<0.05). These findings suggest that different HDM components are associated with various types of allergic diseases. Individualized immunotherapy based on HDM components can enhance treatment efficacy. Chen et al 32 . demonstrated that HDM immunotherapy primarily elicited an immune response characterized by the production of IgG against Der p 1 and Der p 2. Consequently, HDM immunotherapy is effective for patients sensitized exclusively to Der p 1 and Der p 2, but may be ineffective for those sensitized to other dust mite components, such as Der p 5, Der p 7, Der p 21, and Der p 23. This observation may explain why patients with AD experience less favorable outcomes with AIT compared to those with AR and AA. Therefore, it is essential to clarify the role of component detection in the context of AIT. Our study has several limitations. Firstly, the limited number of cases may introduce potential bias. Additionally, our study only included subjects who were positive for Der p-sIgE, excluding those who were positive for Der f-IgE; thus, data from this subset of patients may have been overlooked. Consequently, further studies are required to validate our findings. Nonetheless, our study has contributed to the understanding of HDM allergic patterns and their diversity in China, potentially providing a foundation for personalized prevention and treatment strategies for HDM allergies. Conclusion The establishment of a method for serological analysis of three major HDM allergenic components, sIgE based on a LiCA, enhances our understanding of the clinical symptoms associated with allergic diseases. This novel method addresses the limitations of traditional approaches. Additionally, significant variations in the sensitization patterns of mite allergens were observed across different allergic conditions, with Der p 2 being the most frequently recognized component. A greater complexity in allergic symptoms correlated with an increased positive rate of Der p 23. References 1. Falcon RMG, Caoili SEC. Immunologic, genetic, and ecological interplay of factors involved in allergic diseases. Front Allergy, 2023. 4: p. 1215616. 2. 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Der p 23 sensitization in patients with house dust mite respiratory allergy. Eur Ann Allergy Clin Immunol, 2024. 56(2): p. 79-85. 30. Liu Y, Zhao L, Wang J, Guo Y, Wang Y, Zhang L, Wu Z, Zhu M, Yang X, Xu P, et al. Serological analysis of allergic components of house dust mite provides more insight in epidemiological characteristics and clinical symptom development in North China. Front Immunol, 2023. 14: p. 1083755. 31. Yuriev S, Rodinkova V, Mokin V, Varchuk I, Sharikadze O, Marushko Y, Halushko B, Kurchenko A. Molecular sensitization pattern to house dust mites is formed from the first years of life and includes group 1, 2, Der p 23, Der p 5, Der p 7 and Der p 21 allergens. Clin Mol Allergy, 2023. 21(1): p. 1. 32. Chen KW, Zieglmayer P, Zieglmayer R, Lemell P, Horak F, Bunu CP, Valenta R, Vrtala S. Selection of house dust mite-allergic patients by molecular diagnosis may enhance success of specific immunotherapy. J Allergy Clin Immunol, 2019. 143(3): p. 1248-1252 e12. Figure 1. Effects of three types of component-coated carboxy and aldehyde chemical beads on S/N ratios in serum pools characterized by high, medium, and low levels of Der p sIgE antibodies. A. S/N ratio for Der p 1. B. S/N ratio for Der p 2. C. S/N ratio for Der p 23. Figure 2. The CL signal values at 30, 45, and 60 minutes of incubation were assessed for serum samples containing low to high concentrations of three components. A.CL signal values for Der p 1. B.CL signal values for Der p 2. Figure 3. Prevalence of Sensitization to Der p Components in Der p-Positive Patients Figure 4. The Relationship between HDM Components-sIgE and Allergic Symptom Supplementary Material File (table.docx) Download 20.04 KB Information & Authors Information Version history V1 Version 1 30 June 2025 Peer review timeline Published Journal of Immunological Methods Version of Record 1 Jan 2026 Published Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords allergic diseases component-resolved diagnostics house dust mite light-initiated chemiluminescence assay Authors Affiliations Qinqin Liu The Second Hospital of Tianjin Medical University View all articles by this author Yuanmin Sun Tianjin Medical University View all articles by this author Huiqiang Li Tianjin Medical University View all articles by this author Xue-Yan Wang 0000-0003-3908-3862 [email protected] The Second Hospital of Tianjin Medical University View all articles by this author Metrics & Citations Metrics Article Usage 143 views 90 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Qinqin Liu, Yuanmin Sun, Huiqiang Li, et al. Development and Clinical Application of Component-Resolved Diagnostic Using Light-Initiated Chemiluminescence Assay to characterize House Dust Mite Components-Specific IgE. Authorea . 30 June 2025. 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