A tailored reverse transcription loop-mediated isothermal amplification for sensitive and specific detection of serotype A foot-and-mouth disease virus circulating in pool 1 region countries

• DOI: 10.1111/tbed.12971
• Publication Date: 2018-07-28T03:49:33Z
• Authors: Da-Rae Lim; Hye-Ryung Kim; Min-Ji Park; Ha-Gyeong Chae; Bok-Kyung Ku; Jin-Ju Nah; Soyoon Ryoo; Sung-Hwan Wee; Choi-Kyu Park
• Keywords: Primer (cosmetics)

Rapid and accurate diagnosis of foot-and-mouth disease viruses (FMDV) is essential for the prompt control FMD outbreaks. Reverse transcription polymerase chain reaction (RT-PCR) real-time quantitative RT-PCR (qRT-PCR) are used routine FMDV as World Organisation Animal Health-recommended diagnostic assays. However, these PCR-based assays require sophisticated equipment, specialized labour, complicated procedures detection amplified products, making them unsuitable under-equipped laboratories in developing countries. In this study, to overcome shortcomings, a simple, rapid, cost-effective reverse loop-mediated isothermal amplification (RT-LAMP) assay was developed sensitive specific serotype A circulating pool 1 region. The could be completed 40 min at 62°C, results visually detected by naked eye without any additional systems. specifically VP1 gene Sea-97 genotype FMDV, but it did not amplify other viral nucleic acids. limit 102 TCID50 /ml, which 10 times more than comparable sensitivity qRT-PCR. Evaluation using different strain serotypes showed 100% agreement with RT-PCR. Surprisingly, previously reported RT-LAMP detect all eight tested strains FMDVs, due multiple mismatches between primer template sequences, demonstrating that suitable detecting FMDVs 1-region Conversely, newly improved primers can rapidly accurately diagnose from established study specific, sensitive, tool resource-limited