如何避免COVID-19 PCR检测假阴性和假阳性

I. Fevraleva, O. Glinshchikova, T. Makarik, A. Sudarikov
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引用次数: 0

摘要

背景:在有COVID-19临床表现的情况下,高达40%的COVID-19检测结果可能为阴性。假阴性结果的原因可能源于分析的任何步骤:质量差或空拭子,RNA分离不良,测试中逆转录酶或Taq聚合酶失活。方法:采用人ABL1 mRNA扩增控制拭子质量和完整性的PCR方法检测SARS-CoV-2。设计的引物与ABL1基因的cDNA一起工作,而不是基因组DNA。结果:核衣壳、穗和ABL1基因三个信号同时出现,表明感染了Omicron菌株。ABL1基因和核衣壳扩增仅表明非Omicron感染。ABL1扩增的出现仅表明对SARS-CoV-2的真阴性结果。所有其他的变体都是无效的。结论:已开发出一套SARS-CoV-2多重PCR诊断系统,可消除分析各阶段导致假阴性和假阳性结果的错误。这是通过人类RNA的特定引物的存在,控制正确的拭子应用,处理和RT-PCR的所有阶段来完成的。
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How to Avoid False-Negative and False-Positive COVID-19 PCR Testing
Background: Up to 40% of test results for COVID-19 in the presence of clinical manifestations of the disease might be negative. The reason for a false-negative result might originate from any step of the analysis: poor-quality or empty swab, poor RNA isolation, inactivation of reverse transcriptase or Taq polymerase in the test. Methods: Here we describe a PCR approach for SARS-CoV-2 detection with swab quality and integrity controlled by human ABL1 mRNA amplification. Designed primers work with the cDNA of the ABL1 gene, not genomic DNA. Results: The simultaneous appearance of three signals corresponding to the nucleocapsid, spike, and ABL1 gene indicates infection with the Omicron strain. The amplification of ABL1 gene and nucleocapsid only indicate other than Omicron infection. The appearance of ABL1 amplification only indicates a true negative result for SARS-CoV-2. All other variants are null and void. Conclusions: A system has been developed for multiplex PCR diagnostics of SARS-CoV-2, which makes it possible to eliminate errors leading to false-negative and false-positive results at all stages of analysis. This is accomplished by the presence of specific primers for human RNA, controlling proper swab application, handling, and all the stages of RT-PCR.
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来源期刊
Journal of International Translational Medicine
Journal of International Translational Medicine MEDICINE, RESEARCH & EXPERIMENTAL-
自引率
0.00%
发文量
317
审稿时长
8 weeks
期刊介绍: Journal of International Translational Medicine (JITM, ISSN 2227-6394), founded in 2012, is an English academic journal published by Journal of International Translational Medicine Co., Ltd and sponsored by International Fderation of Translational Medicine. JITM is an open access journal freely serving to submit, review, publish, read and download full text and quote. JITM is a quarterly publication with the first issue published in March, 2013, and all articles published in English are compiled and edited by professional graphic designers according to the international compiling and editing standard. All members of the JITM Editorial Board are the famous international specialists in the field of translational medicine who come from twenty different countries and areas such as USA, Britain, France, Germany and so on.
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