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 ORIGINAL ARTICLE
Year : 2014  |  Volume : 32  |  Issue : 2  |  Page : 124-129

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


1 Department of Obstetrics and Gynecology; Department of Endocrinology and Metabolism, Shengjing Affiliated Hospital of China Medical University, Shenyang 110004, China
2 Department of Endocrinology and Metabolism, 1st Affiliated Hospital of China Medical University, Shenyang 110001, China
3 Department of Medical Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University; Liaoning Center for Disease Control and Prevention, Shenyang 110005, China
4 Department of Medical Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China

Correspondence Address:
C An
Department of Medical Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, Shenyang 110001
China
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Source of Support: This work was supported by the National Natural Science Foundation of China (NSFC 81200653) and Natural Science Funds of Liaoning province, China (2011225020 and 20111108)., Conflict of Interest: We declare that we have no potential confl icts of interest relevant to this article.


DOI: 10.4103/0255-0857.129779

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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.






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