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Updated: Aug 16, 2026
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Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
Published on: March 31, 2021
[Determination of SARS-CoV by simplified nested fluorescent RT-PCR]
Zhen-yong Li1, Da-xiao Shao, Zhi-tao Li
1Key Laboratory of Chem-biology, Department of Chemistry, Zhengzhou University, Zhengzhou 450052, China. zhenyongl@sabc.com.cn
Summary
A new nested reverse transcription-polymerase chain reaction (RT-PCR) method accurately detects SARS coronavirus RNA in patient samples. This rapid and sensitive technique aids in early diagnosis of SARS.
Area of Science:
- Molecular Biology
- Virology
Context:
- Severe Acute Respiratory Syndrome (SARS) emerged in 2002, necessitating rapid diagnostic tools.
- Accurate detection of SARS coronavirus (SARS-CoV) RNA is crucial for disease control.
Purpose:
- To develop a simple, rapid, and sensitive nested reverse transcription-polymerase chain reaction (RT-PCR) assay for SARS-CoV RNA detection.
- To optimize PCR parameters and design specific primers and probes for enhanced sensitivity.
Summary:
- A nested RT-PCR method was established using specific primers and probes targeting SARS-CoV genes.
- Optimization of PCR parameters was performed on a PE 7700 thermal cycler.
- Testing of patient serum (33.6% positive) and mouthwash (67.5% positive) showed significant detection rates, with zero positives in healthy individuals.
Impact:
- The developed nested RT-PCR method offers a rapid, efficient, and sensitive approach for the early diagnosis of SARS.
- This assay can aid in timely identification and management of SARS cases, contributing to public health efforts.
