Establishing an internal quality control method for the stable extraction of nucleic acids of severe acute respiratory syndrome coronavirus 2 and RT‐PCR‐based detection
| Autor: | Kouki Ohtsuka, Wataru Ogura, Shota Yonetani, Satoko Yamasaki, Takahiro Okuyama, Takashi Watanabe, Hiroaki Ohnishi, Hiroyuki Miyagi, Kumiko Sekiguchi |
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| Rok vydání: | 2021 |
| Předmět: |
Quality Control
Microbiology (medical) Coronavirus disease 2019 (COVID-19) Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) media_common.quotation_subject Clinical Biochemistry Computational biology quality control substance SARS‐CoV‐2 COVID‐19 Nucleic Acids Humans Immunology and Allergy Medicine Control chart Quality (business) Research Articles Retrospective Studies media_common SARS-CoV-2 business.industry Biochemistry (medical) Extraction (chemistry) Public Health Environmental and Occupational Health COVID-19 Quality control Hematology real‐time RT‐PCR Medical Laboratory Technology Real-time polymerase chain reaction COVID-19 Nucleic Acid Testing Nucleic acid quality control chart business Research Article |
| Zdroj: | Journal of Clinical Laboratory Analysis |
| ISSN: | 1098-2825 0887-8013 |
| DOI: | 10.1002/jcla.23998 |
| Popis: | Background Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), the causative agent of the coronavirus disease 2019 (COVID‐19), is detected using real‐time RT‐PCR. However, there are limitations pertaining to quality control, particularly with respect to establishing quality control measures for extraction of viral nucleic acids. Here, we investigated the quality control measures for the various processes using an extrinsic quality control substance and quality control charts. Methods An extrinsic quality control substance was added to the sample, and then, real‐time RT‐PCR was performed. Samples with negative test results and the corresponding data were analyzed; a quality control chart was created and examined. Results Data analysis and the quality control charts indicated that SARS‐CoV‐2 could be reliably detected using real‐time RT‐PCR, even when different nucleic acid extraction methods were used or when different technicians were employed. Conclusion With the use of quality control substances, it is possible to achieve quality control throughout the process—from nucleic acid extraction to nucleic acid detection—even upon using varying extraction methods. Further, generating quality control charts would guarantee the stable detection of SARS‐CoV‐2. Real‐time RT‐PCR is used in many countries to detect SARS‐CoV‐2; however, there is no standardized method for quality control during RT‐PCR‐based detection. Our study showed that the use of a quality control substance (EAV) and quality control charts ensures internal quality control for the extraction of SARS‐CoV‐2 nucleic acid and the subsequent virus detection. |
| Databáze: | OpenAIRE |
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