A sensitive duplex nanoparticle-assisted PCR assay for identifying porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus from clinical specimens.
In this study, a novel duplex nanoparticle-assisted polymerase chain reaction (nanoPCR) assay was developed to detect porcine epidemic diarrhea virus (PEDV) and porcine transmissible gastroenteritis virus (TGEV). Two pairs of primers were designed based on the conserved region within the N gene of PEDV and TGEV. In a screening of 114 clinical samples from four provinces in China for PEDV and TGEV, 48.2 and 3.5 % of the samples, respectively, tested positive. Under optimized conditions, the duplex nanoPCR assay had a detection limit of 7.6 × 101and 8.5 × 101copies μL-1for PEDV and TGEV, respectively. The sensitivity of the duplex nanoPCR assay was ten times higher than that of a conventional PCR assay. Moreover, no fragments were amplified when the duplex nanoPCR assay was used to test samples containing other porcine viruses. Our results indicate that the duplex nanoPCR assay described here is useful for the rapid detection of PEDV and TGEV and can be applied in clinical diagnosis.
College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China.|Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China.
Yu
Y
Zhu
Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China.
Lin
L
Liang
Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China.
Yakun
Y
Luo
Animal Medical Center DBN Technology Group, Beijing, 100195, China.
Guihua
G
Wang
College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China.
Chunren
C
Wang
College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China.
Yudong
Y
Cui
College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin, 300384, China. aixialucky@163.com.
Xia
X
Ai
College of Animal Science and Technology, HLJ August First Land Reclamation University, Daqing, 163319, China. cuishangjin@caas.cn.|Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Yuanmingyuan West Road 2, Hai Dian District, Beijing, 100193, China. cuishangjin@caas.cn.
Shangjin
S
Cui
D000818:Animals; D053758:Nanoparticles; D016133:Polymerase Chain Reaction / Q000379:methods; D053485:Porcine epidemic diarrhea virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D005760:Transmissible gastroenteritis virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
Clinical diagnosis; Duplex nanoPCR; N gene; PEDV; TGEV