Rapid detection of Enterovirus 71 and Coxsackievirus A16 by reverse transcriptase loop mediated isothermal amplification (RT-LAMP)

Abstract

   Hand, foot and mouth disease (HFMD) is caused by many types of human enterovirus, in which human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the two main etiological agents. HFMD is common in children under 5 years old and EV71 and CVA16 can cause complications such as encephalitis, myositis leading to death. An effective drug or vaccine against EV71 and CVA16 infection is currently not available. For early treatment it is desirable to accurately and quickly determine the pathogen type of HFMD. Current detection methods are mainly based on clinical symptoms, and RT-PCR is the gold standard for diagnosing. However, RT-PCR requires complicated, expensive equipment and specialized lab environments. Meanwhile, an isothermal nucleic acid amplification method called RT-LAMP has all the benefits of RT-PCR in terms of sensitivity, specificity but with easier technology and low cost. In this study, the optimized RT-LAMP method has been able to detect EV71 and CA16 specific sequences with detection limit of 1 fg for EV71 and 10 fg for CA16. The reaction is performed at a single temperature of 60 ⁰C for 35 minutes; the visible results can be determined by means of the reaction's colour, indicating that the method is highly suitable for point-of-care detection.