Interlaboratory assessment of quantification of SARS-CoV-2 RNA by reverse transcription digital PCR
| Autor: | Yunhua Gao, Yong Guo, Yongzhuo Zhang, Lianhua Dong, Lingxiang Zhu, Chunyan Niu, Nuo Wang, Jingyan Luo, Di Wang, Shangjun Wang, Xia Wang, Ying Jiang, Lin Lu |
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| Rok vydání: | 2021 |
| Předmět: |
Coronavirus disease 2019 (COVID-19)
Copy number Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Absolute quantification Pcr assay RNA COVID-19 Biology Biochemistry Virology RT-dPCR Reverse transcriptase Analytical Chemistry Limit of Detection Quantification Humans RNA Viral Digital polymerase chain reaction Laboratories Gene Paper in Forefront |
| Zdroj: | Analytical and Bioanalytical Chemistry |
| ISSN: | 1618-2650 |
| Popis: | Graphical abstract The pandemic of the novel coronavirus disease 2019 (COVID-19) has caused severe harm to the health of people all around the world. Molecular detection of the pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), played a crucial role in the control of the disease. Reverse transcription digital PCR (RT-dPCR) has been developed and used in the detection of SARS-CoV-2 RNA as an absolute quantification method. Here, an interlaboratory assessment of quantification of SARS-CoV-2 RNA was organized by the National Institute of Metrology, China (NIMC), using in vitro transcribed RNA samples, among ten laboratories on six different dPCR platforms. Copy number concentrations of three genes of SARS-CoV-2 were measured by all participants. Consistent results were obtained with dispersion within 2.2-fold and CV% below 23% among different dPCR platforms and laboratories, and Z′ scores of all the reported results being satisfactory. Possible reasons for the dispersion included PCR assays, partition volume, and reverse transcription conditions. This study demonstrated the comparability and applicability of RT-dPCR method for quantification of SARS-CoV-2 RNA and showed the capability of the participating laboratories at SARS-CoV-2 test by RT-dPCR platform. Supplementary Information The online version contains supplementary material available at 10.1007/s00216-021-03680-2. |
| Databáze: | OpenAIRE |
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