Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials & Methods

LAMP primers

Preparation of the different dilutions of SARS-CoV-2 genome RNA standards

Real-time RT-LAMP reaction

Multiplexed real-time RT-LAMP assay

Colorimetric RT-LAMP reaction

Sensitivity of the triple target RT-LAMP assay for SARS-CoV-2

Specificity of the triple target RT-LAMP assay for SARS-CoV-2

Results

LAMP primers design and selection

LAMP primer sets combination and multiplex RT-LAMP reaction

Visual detection of the SARS-CoV-2 RT-LAMP reaction

Sensitivity of the SARS-CoV-2 triple-target RT-LAMP assay

Specificity of the SARS-CoV-2 triple-target RT-LAMP assay

Discussion

Conclusions

Supplemental Information

Raw data

LC96P files can be opened and analyzed with LightCycler® 96 SW 1.1 software (roche.com).

DOI: 10.7717/peerj.14121/supp-1

Blasting results of the LAMP primer sets

DOI: 10.7717/peerj.14121/supp-2

Reaction times optimization of the RT-LAMP reaction

DOI: 10.7717/peerj.14121/supp-3

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Zhu Yang conceived and designed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Nicole Y. Liu performed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Zhiwei Zhu performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Minmin Xiao analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Shuzhi Zhong analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Qiqi Xue performed the experiments, prepared figures and/or tables, and approved the final draft.

Lina Nie performed the experiments, prepared figures and/or tables, and approved the final draft.

Jinhong Zhao conceived and designed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

The raw data is available in the Supplementary Files.

Funding

This research has been supported by grants from Wuhu Science and Technology Plan Project (No. 2020ms3-9, 2021jc2-6 and 2021yf39), Scientific Research Project of Wannan Medical College (No. wyqnyx202004, WK2021Z07 and JXYY202102), Academic Aid Program for Top-Notch Talents in Provincial Universities (No. gxbjZD2020071), Anhui University Natural Science Research Project (No. KJ2020ZD56). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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