Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630

被引:10
作者
Castellanos-Gonzalez, Alejandro [1 ]
Shelite, Thomas R. [1 ]
Lloyd, Nicole [2 ]
Sadiqova, Aygul [1 ]
Ping, Ren [2 ]
Williams-Bouyer, Natalie [2 ]
Melby, Peter C. [1 ]
Travi, Bruno L. [1 ]
机构
[1] Univ Texas Med Branch, Internal Med Dept, Infect Dis Div, Galveston, TX 77555 USA
[2] Univ Texas Med Branch, Pathol Dept, Infect Dis Div, Galveston, TX 77555 USA
关键词
D O I
10.1038/s41598-021-93333-2
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shortage of kits and supplies to purify viral RNA. Therefore, it is urgent to validate alternative nucleic acid isolation methods for SARS-CoV-2. Our results demonstrate that a concentrated viral lysis amplification buffer (vLAB) prepared with the nonionic detergent IGEPAL enables qualitative detection of SARS-CoV-2 by direct Reverse Transcriptase-Polymerase Chain Reaction (dRT-PCR). Furthermore, vLAB was effective in inactivating SARS-CoV-2. Since this method is inexpensive and no RNA purification equipment or additional cDNA synthesis is required, this dRT-PCR with vLAB should be considered as an alternative method for qualitative detection of SARS-CoV-2.
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页数:10
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