Detection of Enterovirus 71 gene from clinical specimens by reverse-transcription loop-mediated isothermal amplification

作者：Wang D; Wang X; Geng Y; An C^*

来源：Indian Journal of Medical Microbiology, 2014, 32(2): 124-129.

DOI：10.4103/0255-0857.129779

• Clinical specimens
• Enterovirus 71
• hand foot and mouth disease
• reverse transcription loop-mediated isothermal amplification

Purpose: The objective of this study was to develop a sensitive, specific and rapid approach to diagnose hand foot and mouth disease (HFMD) for an early treatment by using loop-mediated isothermal amplification (LAMP) technique. Materials and Methods: A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for detecting EV71 virus was developed, the specificity and sensitivity of RT-LAMP was tested, and the clinical specimens was assayed by the RT-LAMP comparing with conventional reverse-transcription polymerase chain reaction (RT-PCR) and real-time PCR. Results: A total of 116 clinical specimens from the suspected HFMD individual were detected with the RT-LAMP. The detection rate for EV71 was 56.89% by RT-LAMP, 41.38% by real-time PCR and 34.48% by RT-PCR. The minimum detection limit of RT-LAMP was 0.01 PFU, both of RT-PCR and real-time PCR was 0.1PFU. Non-cross-reactive amplification with other enteroviruses was detected in the survey reports. Conclusions: The effectiveness of RT-LAMP is higher than RT-PCR and real-time PCR. The protocol is easy to operate and time saving. It was not an expensive instrument, which was needed; it is an applicable method for rapid diagnosis of the disease, especially in resource-poor countries or in developing countries.

• 出版日期2014-6
• 单位辽宁省疾病预防控制中心; 中国医科大学