구.농수식품
- 조회948
경북대학교 수의과대학, 농림축산검역본부
Abstract
Da‐Rae Lim1,2, Hye‐Ryung Kim1, Ha‐Gyeong Chae1,2, Bok‐Kyung Ku2, Jin‐Ju Nah2, Soyoon Ryoo2, Sung‐Hwan Wee2, Changhee Lee3, Young S. Lyoo4, Choi‐Kyu Park1,*
1College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu, Republic of Korea
2Animal and Plant Quarantine Agency, Gimcheon, Republic of Korea
3Animal Virology Laboratory, School of Life Sciences, Kyungpook National University, Daegu, Republic of Korea
4College of Veterinary Medicine, Konkuk University, Seoul, Republic of Korea
*Correspondence : Choi-Kyu Park, College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea
Abstract
Rapid and specific detection of foot‐and‐mouth disease virus (FMDV) is a key factor for promoting prompt control of FMD outbreaks. In this study, a real‐time reverse transcription loop‐mediated isothermal amplification (RRT‐LAMP) assay with high sensitivity, rapidity and reliability was developed using a targeted gene‐specific assimilating probe for real‐time detection of seven FMDV serotypes. Positive assay signals were generated within 15 min for the lowest concentration of a standard RNA sample at 62°C; this was substantially faster than that achieved by the OIE (World Organisation for Animal Health)‐recommended real‐time quantitative reverse transcription polymerase chain reaction (qRT‐PCR) assay. The new assay specifically amplified the 3D gene of all seven FMDV serotypes and did not amplify other viral nucleic acids. The detection limit of the assay was 102 copies/µl which is comparable to that achieved by qRT‐PCR. Furthermore, using clinical samples, the results of the RRT‐LAMP assay were largely in agreement with those from the qRT‐PCR assay with a kappa value (95% confidence interval [CI]) of 0.94 (0.86–1.02). The established RRT‐LAMP assay that features assimilating probes is an advanced molecular diagnostic tool that is easily applicable to a wide range of circumstances and has high potential for use as an on‐site diagnostic assay for rapid, specific, and reliable detection of FMDVs in clinical samples.
논문정보
- Research article
- 2020년 06월 (BRIC 등록일 2020-06-29)
- 국내 연구진
- 구・농수식품
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