Development, Evaluation, and Clinical Application of PRRSV-2 Vaccine-like Real-Time RT-PCR Assays

Overview

Journal Viruses

Publisher MDPI

Specialty Microbiology

Date 2023 Nov 25

PMID 38005917

Authors

Gaurav Rawal

Karen M Krueger

Wannarat Yim-Im

Ganwu Li

Phillip C Gauger

Marcelo N Almeida

Ethan K Aljets

Jianqiang Zhang

Affiliations

Soon will be listed here.

Abstract

In this study, we developed and validated (1) singleplex real-time RT-PCR assays for specific detection of five PRRSV-2 MLV vaccine viruses (Ingelvac MLV, Ingelvac ATP, Fostera, Prime Pac, and Prevacent) and (2) a four-plex real-time RT-PCR assay (IngelvacMLV/Fostera/Prevacent/XIPC) including the internal positive control XIPC for detecting and distinguishing the three most commonly used vaccines in the USA (Prevacent, Ingelvac MLV, and Fostera). The singleplex and 4-plex vaccine-like PCRs and the reference PCR (VetMAX PRRSV NA&EU, Thermo Fisher Scientific, Waltham, MA, USA) did not cross-react with non-PRRSV swine viral and bacterial pathogens. The limits of detection of vaccine-like PCRs ranged from 25 to 50 genomic copies/reactions. The vaccine-like PCRs all had excellent intra-assay and inter-assay repeatability. Based on the testing of 531 clinical samples and in comparison to the reference PCR, the diagnostic sensitivity, specificity, and agreement were in the respective range of 94.67-100%, 100%, and 97.78-100% for singleplex PCRs and 94.94-100%, 100%, and 97.78-100% for the 4-plex PCR, with a C cutoff of 37. In addition, 45 PRRSV-2 isolates representing different genetic lineages/sublineages were tested with the vaccine-like PCRs and the results were verified with sequencing. In summary, the vaccine-like PCRs specifically detect the respective vaccine-like viruses with comparable performances to the reference PCR, and the 4-plex PCR allows to simultaneously detect and differentiate the three most commonly used vaccine viruses in the same sample. PRRSV-2 vaccine-like PCRs provide an additional tool for detecting and characterizing PRRSV-2.

References

Hu J, Zhang C . Porcine reproductive and respiratory syndrome virus vaccines: current status and strategies to a universal vaccine. Transbound Emerg Dis. 2013; 61(2):109-20. DOI: 10.1111/tbed.12016. View

Liu J, Zhou X, Zhai J, Wei C, Dai A, Yang X . Recombination in JXA1-R vaccine and NADC30-like strain of porcine reproductive and respiratory syndrome viruses. Vet Microbiol. 2017; 204:110-120. DOI: 10.1016/j.vetmic.2017.04.017. View

Yim-Im W, Anderson T, Paploski I, VanderWaal K, Gauger P, Krueger K . Refining PRRSV-2 genetic classification based on global ORF5 sequences and investigation of their geographic distributions and temporal changes. Microbiol Spectr. 2023; 11(6):e0291623. PMC: 10848785. DOI: 10.1128/spectrum.02916-23. View

Zhang Z, Zhou L, Ge X, Guo X, Han J, Yang H . Evolutionary analysis of six isolates of porcine reproductive and respiratory syndrome virus from a single pig farm: MLV-evolved and recombinant viruses. Infect Genet Evol. 2018; 66:111-119. DOI: 10.1016/j.meegid.2018.09.024. View

Brinton M, Gulyaeva A, Balasuriya U, Dunowska M, Faaberg K, Goldberg T . ICTV Virus Taxonomy Profile: 2021. J Gen Virol. 2021; 102(8). PMC: 8513641. DOI: 10.1099/jgv.0.001632. View

Lole K, Bollinger R, Paranjape R, Gadkari D, Kulkarni S, Novak N . Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination. J Virol. 1998; 73(1):152-60. PMC: 103818. DOI: 10.1128/JVI.73.1.152-160.1999. View

Wernike K, Hoffmann B, Dauber M, Lange E, Schirrmeier H, Beer M . Detection and typing of highly pathogenic porcine reproductive and respiratory syndrome virus by multiplex real-time rt-PCR. PLoS One. 2012; 7(6):e38251. PMC: 3387184. DOI: 10.1371/journal.pone.0038251. View

Zhou L, Kang R, Zhang Y, Yu J, Xie B, Chen C . Emergence of two novel recombinant porcine reproductive and respiratory syndrome viruses 2 (lineage 3) in Southwestern China. Vet Microbiol. 2019; 232:30-41. DOI: 10.1016/j.vetmic.2019.01.026. View

Kristensen C, Christiansen M, Pedersen K, Larsen L . Production losses five months after outbreak with a recombinant of two PRRSV vaccine strains in 13 Danish sow herds. Porcine Health Manag. 2020; 6:26. PMC: 7537099. DOI: 10.1186/s40813-020-00165-z. View

10.

Gunson R, Collins T, Carman W . Practical experience of high throughput real time PCR in the routine diagnostic virology setting. J Clin Virol. 2006; 35(4):355-67. PMC: 7108255. DOI: 10.1016/j.jcv.2005.12.006. View

11.

Lunney J, Fang Y, Ladinig A, Chen N, Li Y, Rowland B . Porcine Reproductive and Respiratory Syndrome Virus (PRRSV): Pathogenesis and Interaction with the Immune System. Annu Rev Anim Biosci. 2015; 4:129-54. DOI: 10.1146/annurev-animal-022114-111025. View

12.

Yang K, Tian Y, Zhou D, Duan Z, Guo R, Liu Z . A Multiplex RT-PCR Assay to Detect and Discriminate Porcine Reproductive and Respiratory Syndrome Viruses in Clinical Specimens. Viruses. 2017; 9(8). PMC: 5580462. DOI: 10.3390/v9080205. View

13.

Espy M, Uhl J, Sloan L, Buckwalter S, Jones M, Vetter E . Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev. 2006; 19(1):165-256. PMC: 1360278. DOI: 10.1128/CMR.19.1.165-256.2006. View

14.

Pamornchainavakul N, Kikuti M, Paploski I, Makau D, Rovira A, Corzo C . Measuring How Recombination Re-shapes the Evolutionary History of PRRSV-2: A Genome-Based Phylodynamic Analysis of the Emergence of a Novel PRRSV-2 Variant. Front Vet Sci. 2022; 9:846904. PMC: 8990846. DOI: 10.3389/fvets.2022.846904. View

15.

Renukaradhya G, Meng X, Calvert J, Roof M, Lager K . Live porcine reproductive and respiratory syndrome virus vaccines: Current status and future direction. Vaccine. 2015; 33(33):4069-80. DOI: 10.1016/j.vaccine.2015.06.092. View

16.

Yim-Im W, Huang H, Zheng Y, Li G, Rawal G, Gauger P . Characterization of PRRSV in clinical samples and the corresponding cell culture isolates. Transbound Emerg Dis. 2022; 69(5):e3045-e3059. DOI: 10.1111/tbed.14661. View

17.

Wenhui L, Zhongyan W, Guanqun Z, Zhili L, JingYun M, Qingmei X . Complete genome sequence of a novel variant porcine reproductive and respiratory syndrome virus (PRRSV) strain: evidence for recombination between vaccine and wild-type PRRSV strains. J Virol. 2012; 86(17):9543. PMC: 3416112. DOI: 10.1128/JVI.01341-12. View

18.

Li B, Fang L, Xu Z, Liu S, Gao J, Jiang Y . Recombination in vaccine and circulating strains of porcine reproductive and respiratory syndrome viruses. Emerg Infect Dis. 2009; 15(12):2032-5. PMC: 3044526. DOI: 10.3201/eid1512.090390. View

19.

Wang A, Chen Q, Wang L, Madson D, Harmon K, Gauger P . Recombination between Vaccine and Field Strains of Porcine Reproductive and Respiratory Syndrome Virus. Emerg Infect Dis. 2019; 25(12):2335-2337. PMC: 6874241. DOI: 10.3201/eid2512.191111. View

20.

Bian T, Sun Y, Hao M, Zhou L, Ge X, Guo X . A recombinant type 2 porcine reproductive and respiratory syndrome virus between NADC30-like and a MLV-like: Genetic characterization and pathogenicity for piglets. Infect Genet Evol. 2017; 54:279-286. DOI: 10.1016/j.meegid.2017.07.016. View