An improved reverse transcription loop-mediated isothermal amplification assay for sensitive and specific detection of serotype O foot-and-mouth disease virus

作者：Lim Da Rae; Kim Hye Ryung; Park Min Ji; Chae Ha Gyeong; Ku Bok Kyung; Nah Jin Ju; Ryoo So Yoon; Wee Sung Hwan; Park Yu Ri; Jeon Hyo Sung; Kim Ji Jeong; Jeon Bo Young; Lee Hyeong Woo; Yeo Sang Geon; Park Choi Kyu^*

来源：Journal of Virological Methods, 2018, 260: 6-13.

DOI：10.1016/j.jviromet.2018.06.017

• Foot-and-mouth disease virus
• Serotype-specific
• RT-LAMP
• Swarm primer

A sensitive and specific swarm primer-based reverse transcription loop-mediated isothermal amplification (sRTLAMP) assay for the detection of serotype O foot-and-mouth disease virus (FMDV) was developed and evaluated. The assay specifically amplified the VP3 gene of serotype O FMDV, but did not amplify the VP3 gene of other serotype FMDVs or any other viruses. The limit of detection of the assay was 10(2) TCID50/mL or 10(3) RNA copies/mu L, which is 100 times lower than that of the RT-LAMP assay without swarm primers. The new assay is 10 times more sensitive than reverse transcription-polymerase chain reaction (RT-PCR) and is comparable to the sensitivity of real time RT-PCR (qRT-PCR). Evaluation of the assay using different serotypes of FMDV strains showed 100% agreement with the RT-PCR results. The previously reported serotype O FMDV-specific RT-LAMP assay did not detect 20 out of 22 strains of serotype O FMDVs, probably due to multiple mismatches between the primer and template sequences, showing that it is not suitable for detecting the serotype O FMDVs circulating in Pool 1 region countries, including Korea. In contrast, the developed sRT-LAMP assay with improved primers can rapidly and accurately diagnose serotype O FMDVs circulating in Pool 1 region countries and will be a useful alternative to RT-PCR and qRT-PCR.

• 出版日期2018-10