Simultaneous detection of porcine circovirus type 2, classical swine fever virus, porcine parvovirus and porcine reproductive and respiratory syndrome virus in pigs by multiplex polymerase chain reaction

Jiang, Yonghou; Shang, Hanwu; Xu, Hui; Zhu, Lisha; Chen, Weijie; LingYan, Zhao; Li, Fang

doi:10.1016/j.tvjl.2008.11.016

Cited by 36 publications

(14 citation statements)

References 21 publications

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“…Here it should be emphasized that porcine reproductive and respiratory syndrome caused by PRRS virus is the most economically important viral disease in the swine industry worldwide [26]. Jiang et al [12] used multiplex PCR for the detection of PCV2, classical swine fever virus (CSFV), PPV and porcine reproductive and respiratory syndrome virus (PRRS) in seventy-six clinical specimens. PCV2 was identifi ed using primers for ORF1 region in 9.2% of samples and PPV in 3.9% of samples.…”

Section: Discussionmentioning

confidence: 99%

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Molecular Detection of Pseudorabies Virus (PrV), Porcine Parvovirus (PPV) and Porcine Circovirus 2 (PCV2) in Swine in Republic of Montenegro

et al. 2016

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The presence of pseudorabies virus (PrV), porcine parvovirus (PPV) and porcine circovirus 2 (PCV2) was examined in sixty samples (spleen and lymph nodes) and thirty samples of sacral ganglia collected from non-vaccinated swine by virus isolation and polymerase chain reaction (PCR). Using PCR method PrV was detected in three samples, PPV in seven samples and six samples were found positive for PCV2. The phylogenetic analysis of the nucleotide sequences of three PrV isolates identifi ed in this study showed high similarity and signifi cant clustering within the PrV genotype I strains such as Kaplan and Bartha isolated from pigs in Hungary, strain Becker isolated in USA and strain Kolchis isolated in Greece. The nucleotide sequences of two PPV isolates showed high level of similarity with the strain Challenge isolated from pigs in UK, strain Kresse isolated in USA and strains 77 and LZ isolated in China. The phylogenetic analysis of the nucleotide sequences of two PCV2 isolates showed high level of similarity and signifi cant clustering within genotype PCV2b strains such as NIVS-3, NIVS-5 and NIVS-6 isolated in Serbia, strain 3959 isolated in Austria, strain PM165 isolated from pigs in Brasil, and strain XT2008 isolated in China. The results of our study present the molecular characterization of PrV, PPV and PCV2 identifi ed in swine in Republic of Montenegro. Besides that, these results confi rmed that PCR is a very useful method for rapid detection of these viruses in subclinically infected swine.

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Section: Discussionmentioning

confidence: 99%

“…Virus isolation in cell culture is laborious and time consuming and it cannot be achieved for all PPV strains. Other diagnostic techniques, such as conventional PCR, multiplex PCR or real-time PCR are widely used in the diagnosis of porcine parvovirus [11][12][13].…”

Section: Introductionmentioning

confidence: 99%

Molecular Detection of Pseudorabies Virus (PrV), Porcine Parvovirus (PPV) and Porcine Circovirus 2 (PCV2) in Swine in Republic of Montenegro

et al. 2016

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“…By using multiplex PCR systems, several infectious agents can be detected and differentiated simultaneously in a single reaction, reducing costs and efforts as well as the amount of sample material and time required (37,38). A variety of conventional multiplex PCR assays for the detection and differentiation of different swine viruses based on amplicon size have been developed during recent years (39)(40)(41)(42)(43)(44)(45)(46). However, qPCR assays have several advantages compared to conventional PCR, combining a reduced risk of cross-contamination with a high sensitivity and the possibility of semiquantitative analysis.…”

Section: Discussionmentioning

confidence: 99%

Single-Tube Multiplexed Molecular Detection of Endemic Porcine Viruses in Combination with Background Screening for Transboundary Diseases

Wernike

Hoffmann

2013

J Clin Microbiol

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Detection of several pathogens with multiplexed real-time quantitative PCR (qPCR) assays in a one-step setup allows the simultaneous detection of two endemic porcine and four different selected transboundary viruses. Reverse transcription (RT)-qPCR systems for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2), two of the most economically important pathogens of swine worldwide, were combined with a screening system for diseases notifiable to the World Organization of Animal Health, namely, classical and African swine fever, foot-and-mouth disease, and Aujeszky's disease. Background screening was implemented using the identical fluorophore for all four different RT-qPCR assays. The novel multiplex RT-qPCR system was validated with a large panel of different body fluids and tissues from pigs and other animal species. Both reference samples and clinical specimens were used for a complete evaluation. It could be demonstrated that a highly sensitive and specific parallel detection of the different viruses was possible. The assays for the notifiable diseases were even not affected by the simultaneous amplification of very high loads of PRRSV-and PCV2-specific sequences. The novel broad-spectrum multiplex assay allows in a unique form the routine investigation for endemic porcine pathogens with exclusion diagnostics of the most important transboundary diseases in samples from pigs with unspecific clinical signs, such as fever or hemorrhages. The new system could significantly improve early detection of the most important notifiable diseases of swine and could lead to a new approach in syndromic surveillance.

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“…In recent research, they are concerned about using mPCR to differentiate the highly virulent Chinese-type PRRSV (HP-PRRSV), PRRSV, PPV, CSFV, PRV and PCV2 (Jiang et al 2010). They all cause fulminating infectious diseases in China.…”

Section: Introductionmentioning

confidence: 99%

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Hu¹,

Lin²,

Yang³

et al. 2015

Polish Journal of Veterinary Sciences

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In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.

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Simultaneous detection of porcine circovirus type 2, classical swine fever virus, porcine parvovirus and porcine reproductive and respiratory syndrome virus in pigs by multiplex polymerase chain reaction

Cited by 36 publications

References 21 publications

Molecular Detection of Pseudorabies Virus (PrV), Porcine Parvovirus (PPV) and Porcine Circovirus 2 (PCV2) in Swine in Republic of Montenegro

Molecular Detection of Pseudorabies Virus (PrV), Porcine Parvovirus (PPV) and Porcine Circovirus 2 (PCV2) in Swine in Republic of Montenegro

Single-Tube Multiplexed Molecular Detection of Endemic Porcine Viruses in Combination with Background Screening for Transboundary Diseases

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

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