Development of a novel one‐step triplex PCR assay for the simultaneous detection of porcine circovirus type 2, porcine parvovirus and classical swine fever virus in a single tube. (1st August 2022)

• Record Type: Journal Article
• Authors: Rajkhowa, S.; Choudhury, M.; Pegu, S.R.; Sarma, D.K.; Gupta, V.K.
• Is Part Of: Letters in applied microbiology. Volume 75:Number 2(2022)
• Page Start: 338
• Page End: 344
• Publication Date: 2022-08-01
• Subjects: classical swine fever virus -- porcine circovirus type 2 -- porcine parvovirus -- sensitivity -- specificity -- triplex PCR; Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.13732
• Languages: English
• ISSNs: 0266-8254

Abstract

Abstract: Co‐infection of multiple pathogens complicates diagnosis, treatment and preventive measures based on clinical signs. Therefore, reliable diagnostic tool for timely reporting of suspected diseases is very much essential. A novel one‐step triplex PCR assay was developed and evaluated for simultaneous detection of three important viruses namely porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classical swine fever virus (CSFV) involved in reproductive problems in pigs. Each of the three pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses. The multiplex PCR assay was found to be sensitive in detecting at least 300 pg of viral genomic DNA or RNA from a mixture of three viruses in a reaction. No amplification was obtained from other common viruses or pathogens, such as porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), porcine group A rotavirus (PoRVA), Escherichia coli and Staphylococcus aureus thereby indicating that the developed multiplex PCR has high specificity. Because of the sensitivity and specificity, the developed multiplex PCR assay will be a useful tool for clinical diagnosis of mixed infections of DNA and RNA viruses in pigs. Abstract: Significance and Impact of the Study: The study reports the development and evaluation of a novel one‐step triplex‐PCR assay for simultaneous detection of three important viruses namely porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classical swine fever virus (CSFV) involved in reproductive problems in pigs. As reproductive diseases cause significant economic losses in pig industry and under typical conditions of intensive pig production, it is common for pigs to be simultaneously infected with two or more viral pathogens, there is an urgent need for the development of diagnostic for simultaneous detection of several pathogens in a single reaction which will not only save time but also help in better management of diseases. The assay developed in this study will be very useful for identifying viruses in specimens from pigs with multiple infections.