A novel RT-LAMP assay for rapid and simple detection of classical swine fever virus.
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 °C) 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.
Chen
Lei
L
China Institute of Veterinary Drug Control, Beijing, 100081, China.
Fan
Xue-zheng
XZ
Wang
Qin
Q
Xu
Lu
L
Zhao
Qi-zu
QZ
Zhou
Yuan-chen
YC
Liu
Jun
J
Tang
Bo
B
Zou
Xing-qi
XQ
D000818:Animals; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013552:Swine; D013997:Time Factors; D014773:Virology / Q000379:methods*
D012367:RNA, Viral