An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR
This paper outlines a practical method for validating quantitative-qualitative techniques used to detect genetic material through qRT-PCR, specifically focusing on SARS-CoV-2 testing and adhering to ISO/IEC 17025:2018 accreditation standards. Despite the prevalence of quantitative-qualitative screen...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-01-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024171198 |
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| author | Carolina Cardona-Ramírez Cruz Elena Enríquez-Valencia Gina Méndez-Callejas Giovanna Meza Barreto Gabriel Andrés Tafur-Gómez Danny Wilson Sanjuanelo-Corredor |
| author_facet | Carolina Cardona-Ramírez Cruz Elena Enríquez-Valencia Gina Méndez-Callejas Giovanna Meza Barreto Gabriel Andrés Tafur-Gómez Danny Wilson Sanjuanelo-Corredor |
| author_sort | Carolina Cardona-Ramírez |
| collection | DOAJ |
| description | This paper outlines a practical method for validating quantitative-qualitative techniques used to detect genetic material through qRT-PCR, specifically focusing on SARS-CoV-2 testing and adhering to ISO/IEC 17025:2018 accreditation standards. Despite the prevalence of quantitative-qualitative screening in genetic testing, comprehensive validation guidelines remain a notable gap in the field. Such guidelines could be applied to other molecular testing areas that rely on these techniques, particularly those involving sample handling, automated extraction, and amplification processes, which can significantly impact results. This work describes the statistical approaches behind qRT-PCR protocols, followed by a technical characterization profile of the validation process. Modifications to the gold standard method allowed us to establish a technical limit of detection (LOD) of 5,09 copies/reaction at a 95 % confidence interval. |
| format | Article |
| id | doaj-art-dfac4a045828435493cd4eadd9489798 |
| institution | Kabale University |
| issn | 2405-8440 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj-art-dfac4a045828435493cd4eadd94897982025-01-17T04:50:06ZengElsevierHeliyon2405-84402025-01-01111e41088An integrated methodological framework for the validation and verification of clinical testing by qRT-PCRCarolina Cardona-Ramírez0Cruz Elena Enríquez-Valencia1Gina Méndez-Callejas2Giovanna Meza Barreto3Gabriel Andrés Tafur-Gómez4Danny Wilson Sanjuanelo-Corredor5Grupo de Investigaciones Biomédicas y de Genética Humana Aplicada GIBGA, Facultad de Ciencias de la Salud, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, Colombia; Corresponding author. U.D.C.A, Calle 222 # 55 - 37, Bogotá, Colombia.Grupo de Investigación Ciencia Animal, Facultad de Ciencias Agropecuarias, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, ColombiaGrupo de Investigaciones Biomédicas y de Genética Humana Aplicada GIBGA, Facultad de Ciencias de la Salud, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, Colombia; Corresponding author. U.D.C.A, Calle 222 # 55 - 37, Bogotá, Colombia.Centro de Investigación y Diagnóstico Molecular BSL-3, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, ColombiaGrupo de Investigación Ciencia Animal, Facultad de Ciencias Agropecuarias, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, ColombiaDepartamento de Ciencias Exactas y Naturales, Universidad de Ciencias Aplicadas y Ambientales U.D.C.A, Bogotá, ColombiaThis paper outlines a practical method for validating quantitative-qualitative techniques used to detect genetic material through qRT-PCR, specifically focusing on SARS-CoV-2 testing and adhering to ISO/IEC 17025:2018 accreditation standards. Despite the prevalence of quantitative-qualitative screening in genetic testing, comprehensive validation guidelines remain a notable gap in the field. Such guidelines could be applied to other molecular testing areas that rely on these techniques, particularly those involving sample handling, automated extraction, and amplification processes, which can significantly impact results. This work describes the statistical approaches behind qRT-PCR protocols, followed by a technical characterization profile of the validation process. Modifications to the gold standard method allowed us to establish a technical limit of detection (LOD) of 5,09 copies/reaction at a 95 % confidence interval.http://www.sciencedirect.com/science/article/pii/S2405844024171198AccuracyReliabilityPrecisionSpecificity, SensitivityqRT-PCRISO/IEC 17025 |
| spellingShingle | Carolina Cardona-Ramírez Cruz Elena Enríquez-Valencia Gina Méndez-Callejas Giovanna Meza Barreto Gabriel Andrés Tafur-Gómez Danny Wilson Sanjuanelo-Corredor An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR Heliyon Accuracy Reliability Precision Specificity, Sensitivity qRT-PCR ISO/IEC 17025 |
| title | An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR |
| title_full | An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR |
| title_fullStr | An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR |
| title_full_unstemmed | An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR |
| title_short | An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR |
| title_sort | integrated methodological framework for the validation and verification of clinical testing by qrt pcr |
| topic | Accuracy Reliability Precision Specificity, Sensitivity qRT-PCR ISO/IEC 17025 |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024171198 |
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