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. 2022 Mar 21;18(1):110.
doi: 10.1186/s12917-022-03196-6.

Detection of porcine parainfluenza virus type-1 antibody in swine serum using whole-virus ELISA, indirect fluorescence antibody and virus neutralizing assays

Michael Welch  1 , Karen Krueger  1 , Jianqiang Zhang  1 , Pablo Piñeyro  1 , Ronaldo Magtoto  1 , Chong Wang  1   2 , Luis Giménez-Lirola  1 , Erin Strait  3   4 , Mark Mogler  3 , Phillip Gauger  5
Affiliations

Detection of porcine parainfluenza virus type-1 antibody in swine serum using whole-virus ELISA, indirect fluorescence antibody and virus neutralizing assays

Michael Welch et al. BMC Vet Res. .

Abstract

Background: Porcine parainfluenza virus 1 (PPIV-1) is a respiratory virus in the family Paramyxoviridae and genus Respirovirus. It is closely related to bovine parainfluenza virus 3, human parainfluenza virus 1, and Sendai virus. Recent reports suggest PPIV-1 is widespread in swine herds in the United States and abroad. However, seroprevalence studies and the ability to evaluate cross neutralization between heterologous strains is not possible without validated antibody assays. This study describes the development of an indirect fluorescence antibody (IFA) assay, a whole virus enzyme-linked immunosorbent assay (wv-ELISA) and a serum virus neutralization (SVN) assay for the detection of PPIV-1 antibodies using 521 serum samples collected from three longitudinal studies and two different challenge strains in swine.

Results: The area under the curve (AUC) of the wv-ELISA (95% CI, 0.93-0.98) was significantly higher (p = 0.03) compared to the IFA (95% CI, 0.90-0.96). However, no significant difference was observed between the IFA and wv-ELISA when compared to the SVN (95% CI, 0.92-0.97). All three assays demonstrated relatively uniform results at a 99% true negative rate, with only 11 disagreements observed between the IFA, wv-ELISA and SVN.

Conclusions: All three serology assays detected PPIV-1 antibody in swine serum of known status that was collected from experimental studies. The SVN detected seroconversion earlier compared to the IFA and the wv-ELISA. Both the wv-ELISA and the SVN had similar diagnostic performance, while the IFA was not as sensitive as the wv-ELISA. All three assays are considered valid for routine diagnostic use. These assays will be important for future studies to screen seronegative swine for research, determine PPIV-1 seroprevalence, and to evaluate vaccine efficacy against PPIV-1 under experimental and field conditions.

Keywords: Enzyme linked immunosorbent assay; Indirect fluorescence antibody; Porcine parainfluenza virus 1; Serology; Serum virus neutralization; Validation.

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Conflict of interest statement

Dr. Mark Mogler and Dr. Erin Strait are/were employees of Merck Animal Health, Ames, IA, USA, which provided funding for study C. The additional co-authors declare no competing nor conflict of interest.

Figures

Fig. 1
Fig. 1
Comparative receiver operating characteristic (ROC) analysis. Area under the curve (AUC) was compared between assays. The graph demonstrates the wv-ELISA AUC was significantly increased relative to the IFA (0.96 vs 0.93). However, no significant difference was observed between the SVN and wv-ELISA (0.95 vs 0.96) or SVN and IFA (0.93 vs 0.95)
Fig. 2
Fig. 2
Correlation of antibody concentration among positive samples and percent agreement at a 99% true negative rate. A IFA titer and SVN titer, B IFA titer and wv-ELISA S/P ratio and C wv-ELISA S/P ratio and SVN titer. Figure 2A demonstrated the highest correlation (r = 0.93) between IFA and SVN assays. Similarly, Fig. 2B demonstrated a strong correlation (r = 0.79). The correlation between SVN and wv-ELISA in 2C was lowest at 0.69. SVN: serum virus neutralization, IFA: indirect fluorescence assay, wv-ELISA: whole virus enzyme linked immunosorbent assay
See this image and copyright information in PMC

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