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. 2021 Jun;93(6):3439-3445.
doi: 10.1002/jmv.26736. Epub 2020 Dec 29.

Performance evaluation of antibody tests for detecting infant respiratory syncytial virus infection

Samadhan J Jadhao  1 , Binh Ha  1 , Courtney McCracken  1 , Tebeb Gebretsadik  2 , Christian Rosas-Salazar  2 , James Chappell  2 , Suman Das  2 , Tina Hartert  2 , Larry J Anderson  1
Affiliations

Performance evaluation of antibody tests for detecting infant respiratory syncytial virus infection

Samadhan J Jadhao et al. J Med Virol. 2021 Jun.

Abstract

Respiratory syncytial virus (RSV) infection is a major cause of respiratory tract disease in young children and throughout life. Infant infection is also associated with later respiratory morbidity including asthma. With a prospective birth cohort study of RSV and asthma, we evaluated the performance of an RSV antibody enzyme-linked immunoassay (EIA) for detecting prior infant RSV infection. Infant RSV infection was determined by biweekly respiratory illness surveillance plus RSV polymerase chain reaction (PCR) testing in their first RSV season and serum RSV antibodies after the season at approximately 1 year of age. RSV antibodies were detected by RSV A and B lysate EIA. Antibody and PCR results on 1707 children included 327 RSV PCR positive (PCR+) and 1380 not RSV+. Of 327 PCR+ children, 314 (96%) were lysate EIA positive and 583 out of 1380 (42%) children not PCR+ were positive. We compared the lysate EIA to RSV F, group A G (Ga), and group B G (Gb) protein antibody EIAs in a subset of 226 sera, 118 PCR+ children (97 group A and 21 group B) and 108 not PCR+. In this subset, 117 out of 118 (99%) RSV PCR+ children were positive by both the F and lysate EIAs and 103 out of 118 (87%) were positive by the Ga and/or Gb EIAs. Comparison of the two G EIAs indicated the infecting group correctly in 100 out of 118 (86%) and incorrectly in 1 out of 118 (1%). The lysate and F EIAs are sensitive for detecting infant infection and the two G EIAs can indicate the group of an earlier primary infection.

Keywords: antibodies; infant; infection; respiratory syncytial virus.

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Conflict of interest statement

CONFLICT OF INTERESTS

The authors do not have conflicts related to the contents of this manuscript.

Figures

FIGURE 1
FIGURE 1
Western blot analysis of the supernatant from 293 cells expressing secreted forms of the respiratory syncytial virus (RSV) F, group A G protein (Ga), and group B G protein (Gb). M indicates lane with molecular weight markers. Ctrl indicates lane from 293 cells transfected with empty plasmid. Anti-6X His Mab is an anti-hexahistidine tag monoclonal antibody. Anti-G 3D3 Mab is a human Mab from Trellis Bioscience. Anti-F Motavizumab is a human Mab from MedImmune
FIGURE 2
FIGURE 2
Examples of reference standard curves used to estimate specimen titers for the lysate, F, Ga, and Gb enzyme-linked immunoassays (EIAs). The curves were generated from serial two-fold dilutions of the BEI high titer RSV serum (BEI NR 4021). P is the mean absorbance for three wells against the respective antigen and N is the mean absorbance for three wells against the corresponding negative control antigen. RSV, respiratory syncytial virus
FIGURE 3
FIGURE 3
Distribution of RSV antibody titers for 314 children who were RSV PCR+ and 583 who were not RSV PCR+. A titer >200 is consider positive for RSV antibodies. Sera were collected between the child’s first and second RSV season at approximately 1 year of age. RSV antibodies were detected by an EIA with tissue culture lysate from group A and group B infected cells as the antigen. The titer was estimated from a standard curve. Note the marked difference in number of specimens with low antibody titers, for example, less than 500, between children with RSV PCR+ infection (RSV PCR+) and children with no RSV PCR+ infection (Not RSV PCR+). EIA, enzyme-linked immunoassay; PCR, polymerase chain reaction; RSV, respiratory syncytial virus
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