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利用混合检测优化发展中国家对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的逆转录聚合酶链反应(RT-PCR)检测

Optimizing RT-PCR detection of SARS-CoV-2 for developing countries using pool testing.

作者信息

Farfan Mauricio J, Torres Juan P, O'Ryan Miguel, Olivares Mauricio, Gallardo Pablo, Lastra Jorge, Salas Carolina

机构信息

Hospital Dr. Luis Calvo Mackenna, Santiago, Chile.

Departamento de Pediatría y Cirugía Infantil Oriente, Hospital Dr. Luis Calvo Mackenna, Facultad de Medicina, Universidad de Chile, Santiago, Chile.

出版信息

Rev Chilena Infectol. 2020 Jun;37(3):276-280. doi: 10.4067/s0716-10182020000300276.

Abstract

The global shortage of reagents and kits for nucleic acid extraction and molecular detection of SARS-CoV-2 requires new cost-effective strategies for the diagnosis of suspected COVID-19 cases, especially in countries that need to increase detection capacity. Pooled nucleic acid testing has been extensively used as a cost-effective strategy for HIV, HepB, HepC and influenza. Also, protocols dispensing of RNA extraction appears as an attractive option for detection of SARS-CoV-2. In this study, we found that pooling of 5 samples showed that CT variations were in the range of 1.0-4,5 units, with less likelihood of a false negative result. Results of the sample without nucleic acid ex-traction, was unsatisfactory, with a significant increase in CT values, and thus for risk of a false negative result. In conclusion, pooling nasopharyngeal samples with both automated and manual extraction proved reliable, and thus a potential efficient alternative for the diagnosis of suspected COVID-19 in developing countries.

摘要

全球范围内严重缺乏用于严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核酸提取和分子检测的试剂与试剂盒,这就需要新的具有成本效益的策略来诊断疑似新型冠状病毒肺炎(COVID-19)病例,尤其是在那些需要提高检测能力的国家。混合核酸检测已被广泛用作一种具有成本效益的策略,用于检测艾滋病毒、乙肝病毒、丙肝病毒和流感。此外,省略RNA提取的方案似乎是检测SARS-CoV-2的一个有吸引力的选择。在本研究中,我们发现将5个样本混合后,CT值变化范围为1.0 - 4.5个单位,假阴性结果的可能性较小。未进行核酸提取的样本结果并不理想,CT值显著升高,因此存在假阴性结果的风险。总之,无论是采用自动提取还是手动提取方式,将鼻咽样本混合检测都被证明是可靠的,因此有可能成为发展中国家诊断疑似COVID-19的一种高效替代方法。

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