Foot-and-Mouth Disease Virus Laboratory, Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, India.
J Virol Methods. 2013 Jan;187(1):195-202. doi: 10.1016/j.jviromet.2012.08.015. Epub 2012 Aug 30.
A one-step, real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) for rapid detection and serotyping of Indian foot-and-mouth disease virus (FMDV) is described. The RT-LAMP assay was found to be 10(3)-10(5) fold more sensitive in comparison with RT-PCR, with a detection limit ranging from 10(-3) to 10(-5) TCID(50) of virus samples of all three serotypes. The RT-LAMP assay and qRT-PCR could detect 100 percent of clinical samples of three serotypes, whereas the RT-PCR detected 69.7% of type O, 58.1% of type A and 60.0% of Asia 1 samples. The qRT-PCR has the same sensitivity as the RT-LAMP. The assay conditions with absence of cross reactivity within the three serotypes of FMDV and FMDV negative samples were established. The RT-LAMP assay could detect 100% of samples stored in FTA(®) cards. In comparison with the performance of the RT-PCR; the RT-LAMP appears to be more sensitive, rapid and specific, with the potential for use as a point-of-care (POC) test, especially in developing countries. The use of FTA(®) cards for the preservation of RNA samples coupled with the RT-LAMP assay for the identification of serotypes may help in achieving improved FMDV serotype identification both in the field and in the laboratory.
一种一步法、实时逆转录环介导等温扩增检测方法（RT-LAMP），用于快速检测和定型印度口蹄疫病毒（FMDV）。与 RT-PCR 相比，RT-LAMP 检测法的灵敏度高 10（3）-10（5）倍，病毒样本的检测限范围为 10（-3）-10（-5）TCID（50），三种血清型均适用。RT-LAMP 检测法和 qRT-PCR 可检测到三种血清型的 100%临床样本，而 RT-PCR 仅能检测到 O 型的 69.7%、A型的 58.1%和亚洲 1 型的 60.0%。qRT-PCR 的灵敏度与 RT-LAMP 相同。建立了 RT-LAMP 检测法在三种 FMDV 血清型和 FMDV 阴性样本之间无交叉反应的条件。RT-LAMP 检测法可检测到 100%储存在 FTA（®）卡中的样本。与 RT-PCR 相比，RT-LAMP 具有更高的灵敏度、更快的速度和更高的特异性，有望成为一种即时检测（POC）试验，尤其是在发展中国家。FTA（®）卡用于保存 RNA 样本，结合 RT-LAMP 检测法用于鉴定血清型，有助于提高现场和实验室的 FMDV 血清型鉴定水平。
