[Establishment and application of an indirect ELISA method for detecting respiratory syncytial virus pre-fusion F protein-specific IgG antibodies].

Authors

Y T Wang, X B Liu, H W Xu, F Huang, H Xie

Published in

Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine]. Volume 60. Issue 10. Pages 1622-1629. Oct 06, 2026.

Abstract

Objective: To establish an indirect ELISA method for detecting IgG antibodies specific to the respiratory syncytial virus (RSV) pre-fusion F protein (pre-F), and to conduct systematic validation and application analysis. Methods: The gene sequence of the current circulating RSV strain was obtained through the Beijing respiratory multi-pathogen surveillance system, and the pre-F protein was synthesized as the coating antigen. Key reaction conditions were optimized using a checkerboard titration assay to establish the RSV pre-F ELISA method. Serum samples were randomly selected from healthy community populations and RSV infection convalescent cases. Using plaque reduction neutralization test (PRNT) results as the reference standard, the established method was evaluated for application performance, including sensitivity, specificity, and repeatability, followed by practical application assessment. The assay was also compared in parallel with two commercial ELISA kits, designated kits A and B. Results: The optimal reaction conditions were determined as follows: antigen coating concentration of 5 ng/well, and secondary antibody dilution of 1∶20 000. The cut-off value for positivity was defined as 0.315. The method demonstrated a sensitivity of 98.02%, specificity of 100%, overall agreement of 98.25%, and a Kappa value of 0.92 (P<0.001). Both intra-and inter-assay coefficients of variation were below 5.0%, indicating good repeatability. No significant cross-reactivity was observed with serum samples positive for other common respiratory viruses. Conclusion: This study established an indirect ELISA method for detecting RSV pre-F specific IgG antibodies. The method provides a reliable tool for RSV sero-epidemiological studies and vaccine immunization efficacy evaluation.

PMID:
42893911
Bibliographic data and abstract were imported from PubMed on 19 Oct 2026.

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