Detection of enterovirus 71 gene from clinical specimens by reverse-transcription loop-mediated isothermal amplification.
OBJECTIVE
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.


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.
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D
Wang
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X
Wang
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Y
Geng
Department of Medical Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China.
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An
D002648:Child; D002675:Child, Preschool; D029821:Enterovirus A, Human / Q000235:genetics*; D005260:Female; D006232:Hand, Foot and Mouth Disease / Q000821:virology; D006801:Humans; D007223:Infant; D008297:Male; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction