Reliability of RT-qPCR Pooling Method for COVID-19 Detection in Various Cycle Threshold Values

Althea Medical Journal Pub Date : 2023-06-30 DOI:10.15850/amj.v10n2.2940

Muhammad Fauzan Alif Radjawali, M. Jihadah, L. Chaidir

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Abstract

Background: Reverse Transcriptase Quantitative Real-time Polymerase Chain Reaction (RT-qPCR) is a standard method to detect SARS-CoV-2, the cause of COVID-19 disease, albeit expensive for some laboratory settings. The pooling test is widely used for large-scale screening to speed up the turn-around time and reduce the cost of the RT-qPCR. However, the pooling test involves mixing a certain number of specimens which theoretically increases the possibility of false-negative results. This study aimed to evaluate the accuracy of the pooling test compared with the non-pooling test in different Ct values as a surrogate for viral load.Methods: RT-qPCR was performed in three groups of samples: non-pooling (individual samples), pooling of 5 samples and 11 samples, with various ranges of Ct value in the respective group: x<25 (n=4); 25

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RT-qPCR合并法在不同周期阈值下检测新冠肺炎的可靠性

背景：逆转录酶定量实时聚合酶链反应（RT-qPCR）是检测导致新冠肺炎疾病的SARS-CoV-2的标准方法，尽管在某些实验室环境中成本高昂。池化检测被广泛用于大规模筛查，以加快周转时间并降低RT-qPCR的成本。然而，汇集测试涉及混合一定数量的样本，这在理论上增加了假阴性结果的可能性。本研究旨在评估在不同Ct值下，作为病毒载量替代品的合并试验与非合并试验的准确性。方法：对三组样本进行RT-qPCR：非汇集（单个样本）、汇集5个样本和11个样本，各组的Ct值范围不同：x<25（n=4）；25

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