COVID-19疫情的爆发使各种快速检测方法得以建立和应用。建立一种快速、准确的肠道病毒检测方法尤为重要,这将有利于临床诊断,疫情防控,和及时的可追溯性。通过建立超快速逆转录-聚合酶链反应(RT-PCR)设备,本研究旨在评价基于超快实时荧光RT-PCR技术的肠道病毒核酸检测方法的敏感性和特异性。共抽取61例,然后被运输和保存。核酸提取后,用广州大安基因公司生产的肠道病毒核酸检测试剂盒和本研究建立的超快速RT-PCR设备系统检测同一样品的核酸。ABI7500Fast和Ahram生物系统S1快速设备用于扩增检测。如果样品在FAM通道中具有S形扩增曲线并且Ct值≤40.00,则结果为阳性。敏感性,精度,验证了检测方法的准确性。本研究建立了一种新的检测方法,可在24min内实现肠道病毒核酸的检测。该方法的灵敏度检测极限为1.0×102拷贝/ml。重复检测的变异系数高,中等,低浓度样品为2.644%,1.674%,和4.281%,分别,具有良好的检测重复性。此外,61例可疑样本中,超快速RT-PCR检测方法阳性29例,与常规荧光RT-PCR方法一致。建立的快速检测方法可以大大缩短提供检测报告的时间,可以大大提高诊断和治疗的效率。
The outbreak of COVID-19 epidemic has enabled the establishment and application of various rapid detection methods. It is particularly important to establish a fast and accurate detection method for enterovirus, which will be beneficial for clinical diagnosis, epidemic prevention and control, and timely traceability. Through establishing an ultra-fast reverse transcription-polymerase chain reaction (RT-PCR) equipment, this
study aimed to evaluate the sensitivity and specificity of the testing method of enterovirus nucleic acids based on ultra-fast real-time fluorescence RT-PCR technology. A total of 61 cases were sampled, which were then transported and preserved. After the nucleic acid extraction, the nucleic acids of the same sample were tested with the enterovirus nucleic acid detection kit produced by Guangzhou Da An Gene Company and the ultra-fast RT-PCR equipment system established in this
study. ABI7500Fast and Ahram biosystems S1 fast equipment were used for amplification detection. If the sample had an S-shaped amplification curve in the FAM channel and the Ct value ≤40.00, the result was positive. The sensitivity, precision, and accuracy of the detection method were then verified. This
study established a novel testing method to achieve enterovirus nucleic acid detection within 24 min. The sensitivity detection limit of the method was 1.0 × 102 copies/ml. The coefficients of variation for repeated detection of the high, medium, and low concentration samples were 2.644%, 1.674%, and 4.281%, respectively, with good detection repeatability. In addition, a total of 29 cases were positive by the ultra-fast RT-PCR detection method in 61 suspected samples, which was consistent with the conventional fluorescent RT-PCR method. The established rapid detection method can greatly shorten the time for providing a detection report, which may greatly improve the efficiency of diagnosis and treatment.