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

ABSTRACT

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.