A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus
10.1007/s12250-010-3043-2
- Author:
Lei CHEN
;
Xuezheng FAN
;
Qin WANG
;
Lu XU
;
Qizu ZHAO
;
Yuanchen ZHOU
;
Jun LIU
;
Bo TANG
;
Xingqi ZOU
- Publication Type:Journal Article
- Keywords:
Classical swine fever virus(CSFV);
Reverse transcription loop-mediated isothermal amplification (RT-LAMP);
Rapid detection
- From:
Virologica Sinica
2010;25(1):59-64
- CountryChina
- Language:Chinese
-
Abstract:
A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV.PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.