Detection of porcine parvovirus by loop-mediated isothermal amplification.
Loop-mediated isothermal amplification is a novel method for rapid amplification of DNA. It has been adopted widely for the detection of virus because of its simplicity, rapidity, and specificity. A loop-mediated isothermal amplification assay was developed for the detection of porcine parvovirus. Four primers specific for six regions of PPV non-structural protein 1 gene were designed with an online software. After amplifying at a constant temperature of 59-65 degrees C by Bst enzyme, a clear result was visible after 2.5% agarose gel electrophoresis. The sensitivity and specificity of this assay were evaluated by comparison with the polymerase chain reaction. The detection limit of the assay was shown to be equivalent to 5 PPV copies/reaction. Due to its specificity and simplicity, the assay should be a useful diagnostic tool for epidemiologic studies of PPV.
Chen
Chang-mu
CM
Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 15001, Heilongjiang, China.
Cui
Shang-jin
SJ
D037881:Aborted Fetus / Q000821:virology*; D000034:Abortion, Veterinary / Q000821:virology; D000818:Animals; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010322:Parvoviridae Infections / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D028323:Parvovirus, Porcine / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000453:epidemiology / Q000821:virology*; D013737:Testis / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics
D017931:DNA Primers; D004279:DNA, Viral; D017361:Viral Nonstructural Proteins