Validation and evaluation of RT-PCR real time in house test to detection of SARS-CoV-2 using specific RdRp gene and GAPDH endogen control

Authors

  • Nancy Rojas-Serrano Laboratorio de Referencia Nacional de Virus Respiratorios, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo https://orcid.org/0000-0002-3194-8043
  • Priscila Lope-Pari Laboratorio de Referencia Nacional de Virus Respiratorios, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo https://orcid.org/0000-0002-7439-4351
  • Maribel Huaringa-Nuñez Laboratorio de Referencia Nacional de Virus Respiratorios, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. tecnólogo médico https://orcid.org/0000-0001-9247-4040
  • Paulo Vitor Marques Simas Laboratorio de Patología Aviar, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima, Perú. Laboratorio de Virología Animal, Instituto de Biología, Universidad de Campinas, Campinas/SP, Brasil. Biólogo https://orcid.org/0000-0003-2241-6915
  • Rosa Palacios-Salvatierra Laboratorio de Referencia Nacional de Virus Respiratorios, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo https://orcid.org/0000-0001-9499-2271
  • Johanna Balbuena-Torres Laboratorio de Referencia Nacional de Virus Respiratorios, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo https://orcid.org/0000-0003-4830-6933
  • Omar Alberto Caceres Rey Laboratorio de Referencia Nacional de Biotecnología y Biología Molecular, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo
  • Carlos Padilla-Rojas Laboratorio de Referencia Nacional de Biotecnología y Biología Molecular, Centro Nacional de Salud Pública, Instituto Nacional de Salud, Lima, Perú. Biólogo https://orcid.org/0000-0002-0562-0431

DOI:

https://doi.org/10.17843/rpmesp.2021.384.7596

Keywords:

RT-PCR SARS-CoV-2 testing, SARS-CoV-2, Molecular Diagnosis, COVID-19

Abstract

The present work validated and evaluated a duplex real-time RT-PCR using specific primers and probes for genes RdRp from SARS-CoV-2 and GAPDH from humans; the latter was used as an endogenous control in all reactions. We evaluated the specificity, the sensitivity, the robustness, the reproducibility, the repeatability, the comparability, and the limit of detection. The predictive positive and negative values (PPV and PNV, respectively) and all the parameters evaluated using our duplex real-time RT-PCR was 100%. The detection limit was 100 copies/μL according to the acceptance criteria established for the validation of this protocol. Our duplex real-time RT-PCR demonstrated to be a good alternative for the diagnosis of COVID-19; in addition, this PCR was used adequately in suspicion of COVID-19, allowing it to control the number of false-negatives.

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Published

2021-12-02

Issue

Section

Brief Report

How to Cite

1.
Rojas-Serrano N, Lope-Pari P, Huaringa-Nuñez M, Marques Simas PV, Palacios-Salvatierra R, Balbuena-Torres J, et al. Validation and evaluation of RT-PCR real time in house test to detection of SARS-CoV-2 using specific RdRp gene and GAPDH endogen control. Rev Peru Med Exp Salud Publica [Internet]. 2021 Dec. 2 [cited 2024 Dec. 13];38(4):595-600. Available from: https://rpmesp.ins.gob.pe/index.php/rpmesp/article/view/7596

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