交叉引物扩增检测SARS-CoV-2病毒的有效性

Luhung Budiailmiawan, R. B. Ristandi, A. Fibriani
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引用次数: 0

摘要

检测SARS-CoV-2病毒的标准分子技术是实时逆转录聚合酶链反应(rRT-PCR)。它需要精密的设备和耗时的取样过程。交叉引物扩增(CPA)是一种在恒温条件下对DNA进行高特异性和高效率扩增的核酸扩增技术。这种技术比rRT-PCR更快,而且不需要生物安全二级(BSL-2)设施。本研究旨在确定以rRT-PCR为金标准的CPA的有效性,并评价其作为检测鼻咽拭子标本中SARS-CoV-2 RNA的分子快速检测方法的性能。本研究是一项描述性诊断试验,使用了2021年1月1日至12月31日在Palabuhan Ratu医院治疗的COVID-19患者拭子鼻咽样本的回顾性数据。在Pelabuhan Ratu实验室和省卫生实验室分别对52份鼻咽样本和rRT-PCR样本进行了CPA检测。进行了效度和相关检验。研究对象以34 ~ 50岁的女性为主。截止tt值为3.25,曲线下面积(AUC) 0.84, p值<0.001。CPA对COVID-19的诊断具有良好的有效性,敏感性为77%,特异性为94%,PPV为96%,NPV为71%。敏感度随ct值<30(82%)和ct值<25(87%)而增加。CPA在新冠肺炎诊断测试中具有良好的效度。CPA可作为快速检测鼻咽拭子标本中SARS-CoV-2病毒RNA的分子检测方法。
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The Validity of Cross Priming Amplification to Detect SARS-CoV-2 Virus
The standard molecular technique to detect the SARS-CoV-2 virus is The Real-Time Reverse-Transcription Polymerase Chain Reaction (rRT-PCR). It requires sophisticated equipment and a time-consuming sample process. The Cross Priming Amplification (CPA) is a nucleic acid amplification technique that amplifies DNA with high specificity and efficiency under constant thermal conditions. This technique is faster than rRT-PCR and doesn't require a biosafety level-2 (BSL-2) facility. The study aimed to determine the validity of CPA with rRT-PCR as a gold standard and to evaluate its performance as molecular rapid tests for detecting SARS-CoV-2 RNA from nasopharyngeal swab specimens. This study was a descriptive diagnostic test by using data retrospectively from swab nasopharyngeal patient samples who were treated at Palabuhan Ratu Hospital with COVID-19 from 01 January to 31 December 2021. The CPA was performed on a total of 52 nasopharyngeal samples at Pelabuhan Ratu Laboratory and rRT-PCR at Provincial Health Laboratory. The validity and correlation tests were performed. The majority of subjects were female between the ages of 34-50 years. The cut-off Tt-value is 3.25, 0.84 Area Under Curve (AUC), with a p-value <0.001. The CPA has good validity for COVID-19 diagnosis with 77% sensitivity, 94% specificity, 96% PPV, and 71% NPV. The sensitivity was increasing with Ct-value <30 (82%) and Ct-value <25 (87%). The CPA had a good validity for the COVID-19 diagnostic test. The CPA could be used as a rapid molecular test for detecting SARS-CoV-2 viral RNA from nasopharyngeal swab specimens.
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