A multiplex PCR for rapid and simultaneous detection of porcine circovirus type 2, porcine parvovirus, porcine pseudorabies virus, and porcine reproductive and respiratory syndrome virus in clinical specimens

Virus Genes ◽  
2009 ◽  
Vol 38 (3) ◽  
pp. 392-397 ◽  
Author(s):  
Fengxiong Yue ◽  
Shangjin Cui ◽  
Chaofan Zhang ◽  
Kyoung-Jin Yoon
Keyword(s):  
Multiplex Pcr ◽  
Pseudorabies Virus ◽  
Simultaneous Detection ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Porcine Parvovirus ◽  
Clinical Specimens ◽  
Porcine Pseudorabies Virus

scholarly journals Simultaneous detection of porcine pseudorabies virus, porcine parvovirus and porcine circovirus type 2 by multiplex real-time PCR and amplicon melting curve analysis using SYBR Green I

2018 ◽  
Vol 63 (No. 8) ◽  
pp. 358-366
Author(s):  
LL Zheng ◽  
XH Jin ◽  
FS Wei ◽  
CQ Wang ◽  
HY Chen ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pseudorabies Virus ◽  
Simultaneous Detection ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Porcine Parvovirus ◽  
Porcine Pseudorabies Virus

Porcine parvovirus, porcine pseudorabies virus and porcine circovirus type 2 can cause reproductive failure in pigs, and swine are often simultaneously infected by combinations of the three viruses. We here report the development of a SYBR Green I-based multiplex real time PCR assay for simultaneous detection of porcine parvovirus, porcine pseudorabies virus and porcine circovirus type 2. Three pairs of specific primers were designed for the porcine parvovirus-VP2, porcine pseudorabies virus-gH and porcine circovirus type 2-ORF2 genes. Viral genomes were identified based on their distinctive melting temperatures in singleplex PCR reactions. The melting temperature was 74.5 °C for the 313 bp amplicon of porcine parvovirus-VP2 gene, 87.5 °C for the 355 bp amplicon of porcine pseudorabies virus-gH gene and 80.5 °C for the 171 bp amplicon of the porcine circovirus type 2-ORF2 gene, respectively. The detection limit of the method ranged from 0.01–0.03 TCID<sub>50</sub>/ml for the three viruses. In addition, porcine parvovirus, porcine pseudorabies virus and porcine circovirus type 2 viral loads were measured in 100 field samples, and the result showed that the concordance between real-time PCR and conventional PCR was 60.42%. The sensitivity and specificity of real-time PCR were 100% and 100%, while those of conventional PCR were 40.83% and 72.22%, respectively.

scholarly journals Epidemiological investigation of porcine circovirus type 2 and its coinfection rate in Shandong province in China from 2015 to 2018

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Zicheng Ma ◽  
Mengda Liu ◽  
Zhaohu Liu ◽  
Fanliang Meng ◽  
Hongyu Wang ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Shandong Province ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Limited Information ◽  
Serum Samples ◽  
Diarrhea Virus

Abstract Background Porcine circovirus type 2 (PCV2) is one of the crucial swine viral pathogens, caused porcine circovirus associated diseases (PCVAD). Shandong province is one of the most important pork producing areas and bears a considerable economic loss due to PCVAD. However, there is limited information on epidemiology and coinfection rate of PCV2 with other critical swine diseases in this area, such as porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), Pseudorabies virus (PRV), and porcine epidemic diarrhea virus (PEDV). Results Overall, 89.59% serum samples and 36.98% tissue samples were positive for PCV2 specified ELISA and PCR positive for PCV2, respectively. The coinfection rates of PCV2 with PRRSV, PRV, CSFV, and PEDV were 26.73%, 18.37%, 13.06%, and 3.47%, respectively. Moreover, genetic characteristic of PCV2 were analyzed based on the cap genes showing that PCV2d is the dominant sub-genotype circulating in the province. Conclusions Our findings reveal that PCV2d, as the dominant strain, is prevailing in pig farms in Shandong province at high levels. There was a high frequency of coinfection of PCV2 and PRRSV.

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