A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus

• Published in: Virologica Sinica Vol. 25; no. 1; pp. 59 - 64
• Main Authors: Chen, Lei, Fan, Xue-zheng, Wang, Qin, Xu, Lu, Zhao, Qi-zu, Zhou, Yuan-chen, Liu, Jun, Tang, Bo, Zou, Xing-qi
• Format: Journal Article
• Language: English; Chinese
• Published: Heidelberg SP Wuhan Institute of Virology, CAS 01.02.2010; China Institute of Yeterinary Drug Control,Beijing,100081,China
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bovine viral diarrhea virus; Classical Swine Fever - diagnosis; Classical Swine Fever - virology; Classical swine fever virus; Classical swine fever virus - isolation & purification; Electrophoresis, Agar Gel; Medical Microbiology; Microbial Genetics and Genomics; Microbiology; Molecular Diagnostic Techniques - methods; Nucleic Acid Amplification Techniques - methods; Oncology; Porcine respiratory and reproductive syndrome virus; RNA, Viral - genetics; RNA, Viral - isolation & purification; Sensitivity and Specificity; Simian immunodeficiency virus; Swine; Time Factors; Virology; Virology - methods; 循环使用; 投资工具; 牛病毒性腹泻; 猪瘟病毒; 琼脂糖凝胶电泳; 病毒检测; 逆转录; Rapid detection; Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Classical swine fever virus (CSFV); Classical swine fever virus(CSFV)
• ISSN: 1674-0769; 1995-820X
• DOI: 10.1007/s12250-010-3043-2

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