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.
China Institute of Veterinary Drug Control, Beijing, 100081, China.
Lei
L
Chen
Xue-zheng
XZ
Fan
Qin
Q
Wang
Lu
L
Xu
Qi-zu
QZ
Zhao
Yuan-chen
YC
Zhou
Jun
J
Liu
Bo
B
Tang
Xing-qi
XQ
Zou
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