Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus.
Porcine epidemic diarrhea virus (PEDV) can cause serious disease and even death in neonatal piglets, resulting in serious damage to the swine industry worldwide. Open reading frame 3 (ORF3) is the only accessory gene in the PEDV genome. Previous studies have indicated that PEDV vaccine strains have a partial deletion in ORF3. In this study, a nanoparticle-assisted polymerase chain reaction (nanoparticle-assisted RT-PCR) assay targeting the ORF3 of PEDV was developed to distinguish PEDV field strains from attenuated strains by using a specific pair of primers. The PCR products of field strains and attenuated strains were 264 bp and 215 bp in length, respectively. The sensitivity and specificity of this assay were also assessed. The nanoparticle-assisted RT-PCR assay was 10-100 times more sensitive than the conventional RT-PCR assay, with no cross-reactions when amplifying porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (RV), and porcine transmissible gastroenteritis virus (TGEV). The nanoparticle-assisted RT-PCR assay we describe here can be used to distinguish field strains from vaccine strains of PEDV, and it shows promise for reducing economic loss due to PEDV infection.
Institute of Animal Science(IAS), Chinese Academy of Agricultural Sciences(CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China.|College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China.
Yu
Y
Zhu
Animal Medical Center DBN Technology Group, Beijing, 100195, China.
Gui-Hua
GH
Wang
College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China. 1016856109@qq.com.
Yu-Dong
YD
Cui
Institute of Animal Science(IAS), Chinese Academy of Agricultural Sciences(CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China. cuishangjin@caas.cn.|College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China. cuishangjin@caas.cn.
Shang-Jin
SJ
Cui
D000818:Animals; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D015967:Gene Expression Regulation, Viral; D009154:Mutation; D053758:Nanoparticles; D053485:Porcine epidemic diarrhea virus / Q000145:classification* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014765:Viral Vaccines
D014764:Viral Proteins; D014765:Viral Vaccines