Establishment and preliminary application of RT-LAMP rapid detection method of entervirus 71 / 国际检验医学杂志

ABSTRACT

Objective To establish a reverse transcription loop-mediated isothermal amplification RT-LAMP rapid detection method of enterovirus 71(EV71) in hand ,foot and mouth disease (HFMD) ,and to evaluate its application.Methods Four types of LAMP primers were designed by using the software needle based on the 6 distinct regions of the specific gene of EV71.The process of amplification was completed in the ordinary thermostatic water container by 65 ℃ for 1200 min.The results of amplification were judged by electrophoresis and naked-eye.Seventy EV71 type intestinal positive specimens were simultaneously detected by RT-LAMP and RT-PCR methods.EV71 type of intestinal virus RNA were made a series.After 10 times of dilution ,the RT-LAMP and RT-PCR methods were used to conduct the detection for comparing their sensitivities.Results The LAMP characteristic ladder bands of EV71 appeared ,then the results could judged by the naked-eye.The detection rate in 100 EV71 samples had no statistical difference between RT-LAMP and RT-PCR methods (P>0.05).The sensitivity (10.0 pg/μL) of RT-LAMP was same to that of RT-PCR method.Conclusion The RT-LAMP detection method for EV71 was established ,which can be used for nucleic acid am-plification in the ordinary thermostatic water container.The preliminary application verifies that this RT-LAMP assay has a certain application prospect.