Development and validation of reverse transcription loop-mediated isothermal amplification for detection of PRRSV.
To establish a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of porcine reproductive and respiratory syndrome virus (PRRSV), four primers specific to six regions of the N gene were designed. After amplification in an isothermal water bath for 1 h, samples containing PRRSV generated the expected ladder-like products while porcine parvovirus, porcine circovirus, classic swine fever virus, pseudorabies virus, and swine testis cells generated no product. The sensitivity and specificity of the RT-LAMP assay were evaluated by comparison with reverse-transcription polymerase chain reaction (RT-PCR) and real-time PCR. Because it is specific and simple, the RT-LAMP assay will be useful for the diagnosis of PRRSV infection.
Chen
Changmu
C
Division of Swine Infectious Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of CAAS, No. 427, Maduan Street, Nangang District, Harbin, Heilongjiang 150001, People's Republic of China.
Cui
Shangjin
S
Zhang
Chaofan
C
Li
Jun
J
Wang
Jinbao
J
D000818:Animals; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D013552:Swine; D013696:Temperature