Open AccessMultiplex RT-PCR assay to differentiate genotypes of porcine reproductive and respiratory syndrome virus in swine
Author(s) -
Phat X. Dinh
Publication year - 2019
Publication title -
the journal of agriculture and development/the journal of agriculture and develoment/nông nghiệp và phát triển
Language(s) - English
Resource type - Journals
eISSN - 2615-949X
pISSN - 2615-9503
DOI - 10.52997/jad.2.06.2019
Subject(s) - porcine reproductive and respiratory syndrome virus , multiplex polymerase chain reaction , virology , biology , multiplex , primer (cosmetics) , genotype , real time polymerase chain reaction , arterivirus , gene , virus , polymerase chain reaction , disease , genetics , medicine , covid-19 , pathology , infectious disease (medical specialty) , chemistry , organic chemistry
Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically important diseases to swine industry worldwide. Due to the heterogeneity of field isolates, accurate detection of the PRRS virus is a diagnostic challenge. Recently, co-infection with NA-PRRSV, EU-PRRSV and HP-PRRSV isolates continuously increases in many countries, resulting in a significant impact on PRRSV diagnostics and disease control on farms. To facilitate rapid diagnosis and reliable discrimination of NA-PRRSV, EU-PRRSV and HP-PRRSV, a multiplex RT-PCR assay was established with three pairs of primers targeting highly conservative regions of nsp2 gene with predicted multiplex RT-PCR products of 364 bp, 161 bp and 259 bp, respectively. The primer pairs were optimized to be highly specific for PRRSV genotypes and were able to detect the target gene at the limit of 102 copies/μL for each gene. Clinical samples were used to evaluate this multiplex RT-PCR in parallel with a commercial real-time RT-PCR kit. Results showed over 95.2% (20/21 samples) agreement between the mRT-PCR and the real-time RT-PCR kit. Hence, it indicated that this multiplex RT-PCR could be useful for rapid and deferential diagnosis of NA-PRRSV, EU-PRRSV and HP-PRRSV in swine farms.