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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.
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; Do, Quyet.
Afiliación
  • Nguyen LT; Department of Occupational Medicine, Vietnam Military Medical University, Hanoi, Vietnam.
  • Nguyen PM; Department of Occupational Medicine, Vietnam Military Medical University, Hanoi, Vietnam.
  • Dinh DV; Department of Epidemiology, Vietnam Military Medical University, Hanoi, Vietnam.
  • Pham HN; Department of Epidemiology, Vietnam Military Medical University, Hanoi, Vietnam.
  • Bui LAT; Institute of Biomedicine, Vietnam-Russia Tropical Center, Hanoi, Vietnam.
  • Vo CV; Institute of Biomedicine, Vietnam-Russia Tropical Center, Hanoi, Vietnam.
  • Nguyen BH; Department of Occupational Medicine, Vietnam Military Medical University, Hanoi, Vietnam.
  • Bui HD; Department of Occupational Medicine, Vietnam Military Medical University, Hanoi, Vietnam.
  • Hoang CX; Department of Occupational Medicine, Vietnam Military Medical University, Hanoi, Vietnam.
  • Ngo NMV; Department of Human Anatomy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Dang TT; Department of Human Anatomy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Do AN; Department of Parasitology, Vietnam Military Medical University, Hanoi, Vietnam.
  • Vu DD; Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Nguyen LT; Hanoi University of Science and Technology, Hanoi, Vietnam.
  • Nguyen MN; Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Dinh THT; Faculty of Biology, National University of Hanoi, Hanoi, Vietnam.
  • Ho SA; Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Hoang LV; Faculty of Biology, National University of Hanoi, Hanoi, Vietnam.
  • Hoang SX; Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam.
  • Do Q; Institute of Biomedicine and Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam.
J Clin Lab Anal ; 36(5): e24355, 2022 May.
Article en En | MEDLINE | ID: mdl-35312118
BACKGROUND: 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. OBJECTIVE: We aimed to establish and validate an in-house real-time RT-PCR for the detection of SARS-CoV-2. METHODOLOGY: 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. RESULTS: 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 Ct values observed between two tests with r2  = 0.84 for the E gene and 0.87 for the N gene. Notably, we assessed on 60 paired saliva and nasopharyngeal samples. The overall agreement was 91.66%, and Kappa value of 0.74 showed a high agreement between two types of samples. When using nasopharyngeal swabs as the reference standard, positive percent agreement, and negative percent agreement were 91.83% and 90.90%, respectively. CONCLUSION: In 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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: J Clin Lab Anal Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2022 Tipo del documento: Article País de afiliación: Vietnam Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: SARS-CoV-2 / COVID-19 Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: J Clin Lab Anal Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2022 Tipo del documento: Article País de afiliación: Vietnam Pais de publicación: Estados Unidos