Rapid detection of porcine parvovirus DNA by sensitive loop-mediated isothermal amplification.
A method of loop-mediated isothermal amplification (LAMP) was employed to develop a rapid and simple detection system for porcine parvovirus (PPV) DNA. The amplification could be finished in 45 min under isothermal condition at 62 degrees C by employing a set of four primers targeting VP2 gene of PPV. LAMP assay showed higher sensitivity than PCR, with a detection limit of 5 copies of PPV genomic DNA per reaction. No cross reactivity was observed from the samples of other related viruses including canine parvovirus, parvovirus B19, porcine circovirus type 1, porcine circovirus type 2 and porcine peudorabies virus. The detection rate of PPV LAMP for 125 clinical samples was 97.6% and appeared higher than that of PCR method. The result indicated the potential usefulness of the technique as a simple, rapid procedure for the detection of PPV.
Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Hao-tai
HT
Chen
Jie
J
Zhang
Sheng-hai
SH
Yang
Li-na
LN
Ma
Yan-ping
YP
Ma
Xiang-tao
XT
Liu
Xue-peng
XP
Cai
Yong-guang
YG
Zhang
Yong-sheng
YS
Liu
D000818:Animals; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D028323:Parvovirus, Porcine / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013696:Temperature; D013997:Time Factors
D004279:DNA, Viral