The direct RT-LAMP assay without RNA extraction demonstrated 90.3% sensitivity and 100% specificity compared to RT-LAMP, and 86.83% sensitivity and 100% specificity compared to qRT-PCR for detecting EV71.
Observational (n=145)
No
Does direct RT-LAMP accurately detect EV71 infection in nasopharyngeal swab specimens compared to RT-LAMP and qRT-PCR?
Direct RT-LAMP without RNA extraction is a highly specific and sensitive method for detecting EV71, showing potential for point-of-care screening.
Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD) worldwide and has been associated with neurological complications which resulted in fatalities during recent outbreak in Asia pacific region. A direct reverse transcription loop-mediated isothermal amplification (direct RT-LAMP) assay using heat-treated samples without RNA extraction was developed and evaluated for the detection of EV71 subgenotype C4 in nasopharyngeal swab specimens. The analytical sensitivity and specificity of the direct RT-LAMP assay were examined. The detection limit of the direct RT-LAMP assays was 1.6 of a 50% tissue culture infective dose (TCID(50)) per reaction and no cross-reaction was observed with control viruses including Cosackievirus A (CVA) viruses (CVA2,4,5,7,9,10,14,16, and 24), Coxsackievirus B (CVB) viruses (CVB1,2,3,4, and 5) or ECHO viruses (ECHO3,6,11, and 19). The direct RT-LAMP assay was evaluated and compared to both RT-LAMP and quantitative real-time PCR (qRT-PCR) in detecting EV71 infection with 145 nasopharyngeal swab specimens. The clinical performance demonstrated the sensitivity and specificity of direct RT-LAMP was reported to be 90.3% and 100% respectively, compared to RT-LAMP, and 86.83% and 100% respectively, compared to qRT-PCR. These data demonstrated that the direct RT-LAMP assay can potentially be developed for the point of care screening of EV71 infection in China.
Nie et al. (Tue,) conducted a observational in Hand, foot, and mouth disease (HFMD) / EV71 infection (n=145). Direct RT-LAMP without RNA extraction vs. RT-LAMP with RNA extraction and qRT-PCR was evaluated on Sensitivity and specificity for detecting EV71 compared to RT-LAMP. The direct RT-LAMP assay without RNA extraction demonstrated 90.3% sensitivity and 100% specificity compared to RT-LAMP, and 86.83% sensitivity and 100% specificity compared to qRT-PCR for detecting EV71.