Rapid and Sensitive Detection of PRRSV by a Reverse Transcription-Loop-mediated Isothermal Amplification Assay
10.1007/s12250-011-3185-x
- Author:
Lei ZHANG
;
Yebing LIU
;
Lei CHEN
;
Jianhuan WANG
;
Yibao NING
- Publication Type:Journal Article
- Keywords:
Reverse transcription loop-mediated isothermal amplification (RT-LAMP);
Porcine reproductive and respiratory syndrome virus (PRRSV);
Clinical diagnosis;
Virus detection
- From:
Virologica Sinica
2011;26(4):252-259
- CountryChina
- Language:Chinese
-
Abstract:
A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV)strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries.
