Implementation of Point-of-care Platforms for Rapid Detection of Porcine Circovirus Type 2

Overview

Journal J Vet Sci

Specialty Veterinary Medicine

Date 2024 Apr 3

PMID 38568829

Authors

Chiao-Hsu Ke

Mao-Yuan Du

Wang-Ju Hsieh

Chiu-Chiao Lin

James Mingjuh Ting

Ming-Tang Chiou

Chao-Nan Lin

Affiliations

Soon will be listed here.

Abstract

Background: Porcine circovirus type 2 (PCV2) infection is ubiquitous around the world. Diagnosis of the porcine circovirus-associated disease requires clinic-pathological elements together with the quantification of viral loads. Furthermore, given pig farms in regions lacking access to sufficient laboratory equipment, developing diagnostic devices with high accuracy, accessibility, and affordability is a necessity.

Objectives: This study aims to investigate two newly developed diagnostic tools that may satisfy these criteria.

Methods: We collected 250 specimens, including 170 PCV2-positive and 80 PCV2-negative samples. The standard diagnosis and cycle threshold (Ct) values were determined by quantitative polymerase chain reaction (qPCR). Then, two point-of-care (POC) diagnostic platforms, convective polymerase chain reaction (cPCR, qualitative assay: positive or negative results are shown) and EZtargex (quantitative assay: Ct values are shown), were examined and analyzed.

Results: The sensitivity and specificity of cPCR were 88.23% and 100%, respectively; the sensitivity and specificity of EZtargex were 87.65% and 100%, respectively. These assays also showed excellent concordance compared with the qPCR assay (κ = 0.828 for cPCR and κ = 0.820 for EZtargex). The statistical analysis showed a great diagnostic power of the EZtargex assay to discriminate between samples with different levels of positivity.

Conclusions: The two point-of-care diagnostic platforms are accurate, rapid, convenient and require little training for PCV2 diagnosis. These POC platforms can discriminate viral loads to predict the clinical status of the animals. The current study provided evidence that these diagnostics were applicable with high sensitivity and specificity in the diagnosis of PCV2 infection in the field.

References

Shuai J, Li X, Chen N, Chen X, Fang W . Characterization and potential use of truncated PCV2 capsid protein and its polyclonal antibody for diagnosis of PCV2 infections. Wei Sheng Wu Xue Bao. 2008; 48(1):85-90. View

Nainys J, Lasickiene R, Petraityte-Burneikiene R, Dabrisius J, Lelesius R, Sereika V . Generation in yeast of recombinant virus-like particles of porcine circovirus type 2 capsid protein and their use for a serologic assay and development of monoclonal antibodies. BMC Biotechnol. 2014; 14:100. PMC: 4265424. DOI: 10.1186/s12896-014-0100-1. View

Otoo J, Schlappi T . REASSURED Multiplex Diagnostics: A Critical Review and Forecast. Biosensors (Basel). 2022; 12(2). PMC: 8869588. DOI: 10.3390/bios12020124. View

Segales J, Sibila M . Revisiting Porcine Circovirus Disease Diagnostic Criteria in the Current Epidemiological Context. Vet Sci. 2022; 9(3). PMC: 8953210. DOI: 10.3390/vetsci9030110. View

Lin C, Chan K, Ooi E, Chiou M, Hoang M, Hsueh P . Animal Coronavirus Diseases: Parallels with COVID-19 in Humans. Viruses. 2021; 13(8). PMC: 8402828. DOI: 10.3390/v13081507. View

Opriessnig T, Meng X, Halbur P . Porcine circovirus type 2 associated disease: update on current terminology, clinical manifestations, pathogenesis, diagnosis, and intervention strategies. J Vet Diagn Invest. 2007; 19(6):591-615. DOI: 10.1177/104063870701900601. View

Wang H, Song J, Shin S, Kim H . Comparison of Multiplex Real-Time PCR and PCR-Reverse Blot Hybridization Assays for the Direct and Rapid Detection of Porcine Circovirus Type 2 Genotypes. Front Vet Sci. 2020; 7:200. PMC: 7204941. DOI: 10.3389/fvets.2020.00200. View

Henriques A, Duarte M, Barros S, Fagulha T, Ramos F, Luis T . Development and validation of a real-time PCR for the detection and quantification of porcine circovirus type 2. Virusdisease. 2018; 29(3):355-361. PMC: 6111954. DOI: 10.1007/s13337-018-0476-y. View

Nawagitgul P, Harms P, Morozov I, Thacker B, Sorden S, Lekcharoensuk C . Modified indirect porcine circovirus (PCV) type 2-based and recombinant capsid protein (ORF2)-based enzyme-linked immunosorbent assays for detection of antibodies to PCV. Clin Diagn Lab Immunol. 2002; 9(1):33-40. PMC: 119869. DOI: 10.1128/cdli.9.1.33-40.2002. View

10.

Chianini F, Majo N, Segales J, Dominguez J, Domingo M . Immunohistochemical characterisation of PCV2 associate lesions in lymphoid and non-lymphoid tissues of pigs with natural postweaning multisystemic wasting syndrome (PMWS). Vet Immunol Immunopathol. 2003; 94(1-2):63-75. PMC: 7119727. DOI: 10.1016/s0165-2427(03)00079-5. View

11.

Karuppannan A, Opriessnig T . Porcine Circovirus Type 2 (PCV2) Vaccines in the Context of Current Molecular Epidemiology. Viruses. 2017; 9(5). PMC: 5454412. DOI: 10.3390/v9050099. View

12.

Caprioli A, McNeilly F, McNair I, Lagan-Tregaskis P, Ellis J, Krakowka S . PCR detection of porcine circovirus type 2 (PCV2) DNA in blood, tonsillar and faecal swabs from experimentally infected pigs. Res Vet Sci. 2006; 81(2):287-92. DOI: 10.1016/j.rvsc.2006.01.001. View

13.

Zhou S, Han S, Shi J, Wu J, Yuan X, Cong X . Loop-mediated isothermal amplification for detection of porcine circovirus type 2. Virol J. 2011; 8:497. PMC: 3245487. DOI: 10.1186/1743-422X-8-497. View

14.

Breitbart M, Delwart E, Rosario K, Segales J, Varsani A, Ictv Report Consortium . ICTV Virus Taxonomy Profile: Circoviridae. J Gen Virol. 2017; 98(8):1997-1998. PMC: 5656780. DOI: 10.1099/jgv.0.000871. View

15.

Zhao S, Lin H, Chen S, Yang M, Yan Q, Wen C . Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction. J Vet Diagn Invest. 2015; 27(6):784-8. DOI: 10.1177/1040638715608358. View

16.

Chen H, Zhang J, Sun D, Chu Y, Cai X, Liu X . Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification. J Virol Methods. 2008; 149(2):264-8. PMC: 7112855. DOI: 10.1016/j.jviromet.2008.01.023. View

17.

Jiang M, Wang A, Sun Y, Li Y, Chen Y, Zhou J . Development of a Gold Nanoparticle-Based Immunochromatographic Strip for Rapid Detection of Porcine Circovirus Type 2. Microbiol Spectr. 2023; 11(4):e0195322. PMC: 10434270. DOI: 10.1128/spectrum.01953-22. View

18.

Segales J . Porcine circovirus type 2 (PCV2) infections: clinical signs, pathology and laboratory diagnosis. Virus Res. 2011; 164(1-2):10-9. DOI: 10.1016/j.virusres.2011.10.007. View

19.

Kristensen C, Baadsgaard N, Toft N . A meta-analysis comparing the effect of PCV2 vaccines on average daily weight gain and mortality rate in pigs from weaning to slaughter. Prev Vet Med. 2011; 98(4):250-8. DOI: 10.1016/j.prevetmed.2010.11.015. View

20.

Kim D, Ha Y, Lee Y, Chae S, Lee K, Han K . Comparative study of in situ hybridization and immunohistochemistry for the detection of porcine circovirus 2 in formalin-fixed, paraffin-embedded tissues. J Vet Med Sci. 2009; 71(7):1001-4. DOI: 10.1292/jvms.71.1001. View