1. Establishment of an in‐house real‐time RT‐PCR assay for the detection of severe acute respiratory syndrome coronavirus 2 using the first World Health Organization international standard in a resource‐limited country
- Author
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Nguyen, Linh Tung, Nguyen, Phuong Minh, Dinh, Duc Viet, Pham, Hung Ngoc, Bui, Lan Anh Thi, Vo, Cuong Viet, Nguyen, Ben Huu, Bui, Hoan Duy, Hoang, Cuong Xuan, Ngo, Nhat Minh Van, Dang, Truong Tien, Do, Anh Ngoc, Vu, Dung Dinh, Nguyen, Linh Thuy, Nguyen, Mai Ngoc, Dinh, Thu Hang Thi, Ho, Son Anh, Hoang, Luong Van, Hoang, Su Xuan, and Do, Quyet
- Subjects
Microbiology (medical) ,Reverse Transcriptase Polymerase Chain Reaction ,SARS-CoV-2 ,Biochemistry (medical) ,Clinical Biochemistry ,Public Health, Environmental and Occupational Health ,COVID-19 ,Hematology ,World Health Organization ,Sensitivity and Specificity ,Medical Laboratory Technology ,Nasopharynx ,Humans ,Immunology and Allergy ,Pandemics - Abstract
The COVID-19 pandemic caused by SARS-CoV-2 remains public health burdens and many unresolved issues worldwide. Molecular assays based on real-time RT-PCR are critical for the detection of SARS-CoV-2 in clinical specimens from patients suspected of COVID-19.We aimed to establish and validate an in-house real-time RT-PCR for the detection of SARS-CoV-2.Primers and probes sets in our in-house real-time RT-PCR assay were designed in conserved regions of the N and E target genes. Optimized multiplex real-time RT-PCR assay was validated using the first WHO International Standard (NIBSC code: 20/146) and evaluated clinical performance.The limit of detection validated using the first WHO International Standard was 159 IU/ml for both E and N target genes. The evaluation of clinical performance on 170 clinical samples showed a positive percent agreement of 100% and the negative percent agreement of 99.08% for both target genes. The Kappa value of 0.99 was an excellent agreement, the strong correlation of CIn the present study, we established and validated an in-house real-time RT-PCR for molecular detection of SARS-CoV-2 in a resource-limited country.
- Published
- 2022