Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA
IntroductionVaccines capable of effectively inducing mucosal immunity, particularly specific IgA antibodies, represent an ideal strategy for preventing infections and the transmission of pathogens such as SARS-CoV-2 and influenza viruses that rapidly replicate in the upper respiratory tract and caus...
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Frontiers Media S.A.
2025-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1571418/full |
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| author | Xuanxuan Zhang Xuanxuan Zhang Jizong Jia Chaoying Hu Chaoying Hu Yulong Fu Guanxing Liu Yajing Li Qian He Qian He Fan Gao Fan Gao Na Li Lina Wang Jianping Chu Henggang Xu Zhihao Fu Zhihao Fu Hui Zhao Hui Zhao Zhenglun Liang Zhenglun Liang Jingxin Li Miao Xu Miao Xu Qunying Mao Qunying Mao |
| author_facet | Xuanxuan Zhang Xuanxuan Zhang Jizong Jia Chaoying Hu Chaoying Hu Yulong Fu Guanxing Liu Yajing Li Qian He Qian He Fan Gao Fan Gao Na Li Lina Wang Jianping Chu Henggang Xu Zhihao Fu Zhihao Fu Hui Zhao Hui Zhao Zhenglun Liang Zhenglun Liang Jingxin Li Miao Xu Miao Xu Qunying Mao Qunying Mao |
| author_sort | Xuanxuan Zhang |
| collection | DOAJ |
| description | IntroductionVaccines capable of effectively inducing mucosal immunity, particularly specific IgA antibodies, represent an ideal strategy for preventing infections and the transmission of pathogens such as SARS-CoV-2 and influenza viruses that rapidly replicate in the upper respiratory tract and cause clinical symptoms. However, a lack of standardized nasal antibody detection and sampling methods has hindered cross-study comparability and vaccine development.MethodsThis study uses SARS-CoV-2 as a model pathogen to standardize nasal antibody detection methods and sampling methods. Following the scientific guidelines (Q14 and Q2(R2)) for analytical procedure development and validation released by the International Council for Harmonization (ICH), an ELISA for nasal SARS-CoV-2 WT-RBD specific IgA detection was established and validated. To compare the sampling methods, nasal samples were collected from five groups using three commonly used nasal sampling methods (M1: nasopharyngeal swab; M2: nasal swab; M3: expanding sponge method). The total IgA and SARS-CoV-2 WT-RBD IgA in clinical samples were detected.ResultsThe first validated ELISA for nasal SARS-CoV-2 WT-RBD specific IgA detection was established through analytical target profiling (ATP), risk assessment, and design of experiment optimization. Systematic validation demonstrated exclusive specificity for the target antigen, with intermediate precision of <17% and relative bias of <±4%, meeting ATP requirements. Analysis of 154 clinical samples demonstrated strong concordance between the novel method and electrochemiluminescence assays, with a concordance correlation coefficient of 0.87 for quantitative results and a kappa coefficient of 0.85 for results above and below the dilution-adjusted limit of quantification (LOQ). Applying this novel method, a clinical comparison revealed that M3 achieved superior performance in terms of the single-day detection rate (above dilution-adjusted LOQ 95.5%), 5-day consecutive detection rate (above dilution-adjusted LOQ 88.9%), and median SARS-CoV-2 WT-RBD IgA concentration (171.2 U/mL), significantly outperforming M1 (68.8%; 48.7%; 28.7 U/mL, p<0.0001) and M2 (88.3%; 77.3%; 93.7 U/mL, p<0.05).ConclusionThis study has established the first standardized nasal detection system. The system can be adapted with appropriate modifications for the clinical evaluation of other respiratory mucosal vaccines, thereby advancing the development of mucosal vaccines. |
| format | Article |
| id | doaj-art-b4db16b9f3cb41aa8676d254b2f8873c |
| institution | OA Journals |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-b4db16b9f3cb41aa8676d254b2f8873c2025-08-20T01:52:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.15714181571418Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgAXuanxuan Zhang0Xuanxuan Zhang1Jizong Jia2Chaoying Hu3Chaoying Hu4Yulong Fu5Guanxing Liu6Yajing Li7Qian He8Qian He9Fan Gao10Fan Gao11Na Li12Lina Wang13Jianping Chu14Henggang Xu15Zhihao Fu16Zhihao Fu17Hui Zhao18Hui Zhao19Zhenglun Liang20Zhenglun Liang21Jingxin Li22Miao Xu23Miao Xu24Qunying Mao25Qunying Mao26State Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaBeijing Wantai Biological Pharmacy Enterprise Co., Ltd, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaSchool of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, ChinaChangchun Institute of Biological Products Co., Ltd., Changchun, ChinaShanghai Institute of Biological Products Co., Ltd., Shanghai, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaBeijing Minhai Biotechnology Co., Ltd., Beijing, ChinaBeijing Wantai Biological Pharmacy Enterprise Co., Ltd, Beijing, ChinaBeijing Wantai Biological Pharmacy Enterprise Co., Ltd, Beijing, ChinaBeijing Wantai Biological Pharmacy Enterprise Co., Ltd, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaJiangsu Provincial Centre for Disease Control and Prevention, Public Health Research Institute of Jiangsu Province, Nanjing, Jiangsu, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaState Key Laboratory of Drug Regulatory Science, Evaluation of Biological Products, Key Laboratory of Research on Quality and Standardization of Biotech Products, Institute of Biological Products, National Institutes for Food and Drug Control, Beijing, ChinaResearch Units of Innovative Vaccine Quality Evaluation and Standardization, Chinese Academy of Medical Sciences, Beijing, ChinaIntroductionVaccines capable of effectively inducing mucosal immunity, particularly specific IgA antibodies, represent an ideal strategy for preventing infections and the transmission of pathogens such as SARS-CoV-2 and influenza viruses that rapidly replicate in the upper respiratory tract and cause clinical symptoms. However, a lack of standardized nasal antibody detection and sampling methods has hindered cross-study comparability and vaccine development.MethodsThis study uses SARS-CoV-2 as a model pathogen to standardize nasal antibody detection methods and sampling methods. Following the scientific guidelines (Q14 and Q2(R2)) for analytical procedure development and validation released by the International Council for Harmonization (ICH), an ELISA for nasal SARS-CoV-2 WT-RBD specific IgA detection was established and validated. To compare the sampling methods, nasal samples were collected from five groups using three commonly used nasal sampling methods (M1: nasopharyngeal swab; M2: nasal swab; M3: expanding sponge method). The total IgA and SARS-CoV-2 WT-RBD IgA in clinical samples were detected.ResultsThe first validated ELISA for nasal SARS-CoV-2 WT-RBD specific IgA detection was established through analytical target profiling (ATP), risk assessment, and design of experiment optimization. Systematic validation demonstrated exclusive specificity for the target antigen, with intermediate precision of <17% and relative bias of <±4%, meeting ATP requirements. Analysis of 154 clinical samples demonstrated strong concordance between the novel method and electrochemiluminescence assays, with a concordance correlation coefficient of 0.87 for quantitative results and a kappa coefficient of 0.85 for results above and below the dilution-adjusted limit of quantification (LOQ). Applying this novel method, a clinical comparison revealed that M3 achieved superior performance in terms of the single-day detection rate (above dilution-adjusted LOQ 95.5%), 5-day consecutive detection rate (above dilution-adjusted LOQ 88.9%), and median SARS-CoV-2 WT-RBD IgA concentration (171.2 U/mL), significantly outperforming M1 (68.8%; 48.7%; 28.7 U/mL, p<0.0001) and M2 (88.3%; 77.3%; 93.7 U/mL, p<0.05).ConclusionThis study has established the first standardized nasal detection system. The system can be adapted with appropriate modifications for the clinical evaluation of other respiratory mucosal vaccines, thereby advancing the development of mucosal vaccines.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1571418/fullnasal antibodynasal sample collectionbinding activityimmune assaySARS-CoV-2 |
| spellingShingle | Xuanxuan Zhang Xuanxuan Zhang Jizong Jia Chaoying Hu Chaoying Hu Yulong Fu Guanxing Liu Yajing Li Qian He Qian He Fan Gao Fan Gao Na Li Lina Wang Jianping Chu Henggang Xu Zhihao Fu Zhihao Fu Hui Zhao Hui Zhao Zhenglun Liang Zhenglun Liang Jingxin Li Miao Xu Miao Xu Qunying Mao Qunying Mao Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA Frontiers in Immunology nasal antibody nasal sample collection binding activity immune assay SARS-CoV-2 |
| title | Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA |
| title_full | Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA |
| title_fullStr | Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA |
| title_full_unstemmed | Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA |
| title_short | Exploring the standardized detection and sampling methods of human nasal SARS-CoV-2 RBD IgA |
| title_sort | exploring the standardized detection and sampling methods of human nasal sars cov 2 rbd iga |
| topic | nasal antibody nasal sample collection binding activity immune assay SARS-CoV-2 |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1571418/full |
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