Development and Use of a Multiplex Real-Time Quantitative Polymerase Chain Reaction Assay for Detection and Differentiation of Porcine Circovirus-2 Genotypes 2a and 2b in an Epidemiological Survey

Journal of Veterinary Diagnostic Investigation - Tập 20 Số 5 - Trang 545-558 - 2008
Carl A. Gagnon1,2, J Castillo1,2, Nedzad Music1, Guy Fontaine2, Josée Harel1,2, Donald Tremblay2
1Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada
2Service de diagnostic, and the Groupe de recherche en pharmacologie animale du Québec, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada.

Tóm tắt

By the end of 2004, the Canadian swine population had experienced a severe increase in the incidence of Porcine circovirus-associated disease (PCVAD), a problem that was associated with the emergence of a new Porcine circovirus-2 genotype (PCV-2b), previously unrecovered in North America. Thus, it became important to develop a diagnostic tool that could differentiate between the old and new circulating genotypes (PCV-2a and PCV-2b, respectively). Consequently, a multiplex real-time quantitative polymerase chain reaction (mrtqPCR) assay that could sensitively and specifically identify and differentiate PCV-2 genotypes was developed. A retrospective epidemiologic survey that used the mrtqPCR assay was performed to determine if cofactors could affect the risk of PCVAD. From 121 PCV-2-positive cases gathered for this study, 4.13%, 92.56%, and 3.31% were positive for PCV-2a, PCV-2b, and both genotypes, respectively. In a data analysis using univariate logistic regressions, the PCVAD-compatible (PCVAD/c) score was significantly associated with the presence of Porcine reproductive and respiratory syndrome virus (PRRSV), PRRSV viral load, PCV-2 viral load, and PCV-2 immunohistochemistry (IHC) results. Polytomous logistic regression analysis revealed that PCVAD/c score was affected by PCV-2 viral load ( P = 0.0161) and IHC ( P = 0.0128), but not by the PRRSV variables ( P > 0.9), which suggests that mrtqPCR in tissue is a reliable alternative to IHC. Logistic regression analyses revealed that PCV-2 increased the odds ratio of isolating 2 major swine pathogens of the respiratory tract, Actinobacillus pleuropneumoniae and Streptococcus suis serotypes 1/2, 1, 2, 3, 4, and 7, which are serotypes commonly associated with clinical diseases.

Từ khóa


Tài liệu tham khảo

10.1177/104063879801000102

10.1093/ije/26.6.1323

10.1016/S0895-4356(98)00066-3

10.1016/j.jviromet.2004.08.014

Carman S, 2008, Can J Vet Res, 72, 259

Carman S, 2006, Can Vet J, 47, 761

10.1016/S1090-0233(03)00182-5

10.1016/j.tvjl.2004.01.012

10.1016/S0042-6822(03)00096-5

10.1006/viro.2002.1733

10.1007/s00705-006-0909-6

10.1016/j.jviromet.2004.10.003

Clark EG: 1997, Post-weaning multisystemic wasting syndrome. In: 28th annual meeting of the American Association of Swine Practitioners, pp. 499–501. AASP, Quebec City, Canada.

D'Allaire S, 2007, Can Vet J, 48, 145

10.1099/vir.0.19536-0

10.2460/javma.230.2.244

10.1016/j.vetmic.2007.09.016

10.1016/j.vetmic.2003.10.008

Ellis J, 1998, Can Vet J, 39, 44

10.1128/JCM.38.7.2494-2503.2000

10.1128/JVI.78.24.13440-13446.2004

10.1007/s00705-002-0943-y

Gagnon CA, 2007, Can Vet J, 48, 811

10.1016/j.vetmic.2007.09.007

10.1128/JVI.72.6.5262-5267.1998

Harding JC: 1997, Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation. In: 28th annual meeting of the American Association of Swine Practitioners, p. 503. AASP, Quebec City, Canada.

10.1016/j.vetmic.2003.10.013

10.1016/j.virusres.2007.10.013

Higgins R, 1999, Diseases of swine, 8, 563

Horlen KP, 2007, J Swine Health Prod, 15, 270, 10.54846/jshap/520

Hosmer DW, 1989, Applied logistic regression

10.1177/104063870401600108

10.1016/S0168-1702(02)00141-7

10.1016/j.jviromet.2006.09.003

10.1128/JVI.80.10.5065-5073.2006

10.1128/JVI.79.13.8262-8274.2005

10.1099/vir.0.82629-0

10.1006/viro.2000.0730

McCullagh P, 1980, J Royal Stat Soc Series B, 42, 109, 10.1111/j.2517-6161.1980.tb01109.x

10.1177/104063870201400203

10.1099/0022-1317-79-9-2171

10.1099/0022-1317-81-9-2281

10.1016/j.virol.2006.07.047

10.1016/j.jviromet.2003.12.007

10.1354/vp.41-6-624

10.1128/JCM.37.12.3917-3924.1999

10.2307/j.ctv18pgnrq

10.1177/104063870201400601

10.1128/CDLI.11.4.736-741.2004

10.1128/JVI.76.7.3232-3239.2002

10.1079/AHR2005106

10.1016/S0378-1135(01)00474-6

10.1177/104063879901100607

10.1038/295064a0

10.1631/jzus.2007.B0162

Thông tin
Thông tin xuất bản

Journal of Veterinary Diagnostic Investigation

Tập 20 Số 5

545-558

Thông tin tác giả