Fast SARS-CoV-2 detection by RT-qPCR in preheated nasopharyngeal swab samples

被引:50
|
作者
Alcoba-Florez, Julia [1 ]
Gonzalez-Montelongo, Rafaela [2 ]
inigo-Campos, Antonio [2 ]
de Artola, Diego Garcia-Martinez [1 ]
Gil-Campesino, Helena [1 ]
Ciuffreda, Laura [4 ]
Valenzuela-Fernandez, Agustin [5 ,6 ]
Flores, Carlos [2 ,7 ]
机构
[1] Hosp Univ NS Candelaria, Serv Microbiol, Santa Cruz De Tenerife 38010, Spain
[2] Inst Tecnol & Energias Renovables, Genom Div, Santa Cruz De Tenerife 38600, Spain
[3] Hosp Univ NS Candelaria, Res Unit, Santa Cruz De Tenerife 38010, Spain
[4] Univ La Laguna, Fac Med, Unidad Farmacol, Lab Inmunol Celular & Viral, San Cristobal De La Lagu 38200, Spain
[5] Inst Salud Carlos III, Red Espanola Invest VIH SIDA RIS RETIC, Madrid 28029, Spain
[6] Inst Salud Carlos III, CIBER Enfermedades Resp, Madrid 28029, Spain
[7] Univ La Laguna, Inst Tecnol Biomed ITB, San Cristobal De La Lagu 38200, Spain
关键词
COVID-19; SARS-CoV-2; diagnosis; sample treatment; RNA extraction; fast protocols;
D O I
10.1016/j.ijid.2020.05.099
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
Objectives: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for highthroughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. Methods: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnap (TM) buffer. Results: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70 degrees C incubation for 10 min. Conclusions: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis. (C) 2020 The Author(s). Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-ncnd/4.0/).
引用
收藏
页码:66 / 68
页数:3
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