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.
A multiplex polymerase chain reaction (PCR) was designed for the simultaneous detection of four viruses involved in reproductive and respiratory failure in pigs: porcine circovirus type 2 (PCV-2), porcine parvovirus (PPV), classical swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses. The sensitivity of the multiplex PCR using purified plasmid constructs containing the specific viral target fragments was 2.58x10(7), 2.64x10(5), 2.66x10(7) and 2.73x10(5) copies for PRRSV, PCV-2, CSFV and PPV, respectively. Using the multiplex PCR, co-infections with these four viruses were identified in 26/76 (34.2%) piglets born from sows with reproductive failure in China. This method is a rapid, sensitive and cost-effective diagnostic tool for the routine surveillance of viral infections in pigs.
Jiang
Yonghou
Y
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China. yonghoujiang@zstu.edu.cn
Shang
Hanwu
H
Xu
Hui
H
Zhu
Liangjun
L
Chen
Weijie
W
Zhao
Lingyan
L
Fang
Li
L
D000818:Animals; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary; D017925:Circovirus / Q000235:genetics / Q000302:isolation & purification; D006691:Classical Swine Fever / Q000175:diagnosis; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary; D028323:Parvovirus, Porcine / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D018571:Sentinel Surveillance / Q000662:veterinary; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary*; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral