tetano
Editor, Senior Moderator
Microb Biotechnol. 2020 Apr 25. doi: 10.1111/1751-7915.13586. [Epub ahead of print]
RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2.
Huang WE[SUP]1,[/SUP][SUP]2[/SUP], Lim B[SUP]2[/SUP], Hsu CC[SUP]2,[/SUP][SUP]3[/SUP], Xiong D[SUP]4[/SUP], Wu W[SUP]4[/SUP], Yu Y[SUP]2,[/SUP][SUP]3[/SUP], Jia H[SUP]1[/SUP], Wang Y[SUP]1[/SUP], Zeng Y[SUP]1[/SUP], Ji M[SUP]1[/SUP], Chang H[SUP]1[/SUP], Zhang X[SUP]4[/SUP], Wang H[SUP]1[/SUP], Cui Z[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP].
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Abstract
The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.
? 2020 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.
PMID:32333644DOI:10.1111/1751-7915.13586
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RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2.
Huang WE[SUP]1,[/SUP][SUP]2[/SUP], Lim B[SUP]2[/SUP], Hsu CC[SUP]2,[/SUP][SUP]3[/SUP], Xiong D[SUP]4[/SUP], Wu W[SUP]4[/SUP], Yu Y[SUP]2,[/SUP][SUP]3[/SUP], Jia H[SUP]1[/SUP], Wang Y[SUP]1[/SUP], Zeng Y[SUP]1[/SUP], Ji M[SUP]1[/SUP], Chang H[SUP]1[/SUP], Zhang X[SUP]4[/SUP], Wang H[SUP]1[/SUP], Cui Z[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP].
Author information
Abstract
The pandemic coronavirus SARS-CoV-2 in the world has caused a large infected population suffering from COVID-19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to achieve the detection of SARS-CoV-2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS-CoV-2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT-LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT-qPCR. In addition, we also show that one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT-LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.
? 2020 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.
PMID:32333644DOI:10.1111/1751-7915.13586
Free full text