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An international, interlaboratory ring trial confirms the feasibility of an extraction-less "direct" RT-qPCR method for reliable detection of SARS-CoV-2 RNA in clinical samples

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dc.contributor.authorMills, Margaret
dc.contributor.authorBruce, Emily
dc.contributor.authorHuang, Meei-Li
dc.contributor.authorCrothers, Jessica
dc.contributor.authorHyrien, Ollivier
dc.contributor.authorOura, Christopher
dc.contributor.authorBlake, Lemar
dc.contributor.authorBrown, Arianne
dc.contributor.authorHester, Susan
dc.contributor.authorWehmas, Leah
dc.contributor.authorMari, Bernard
dc.contributor.authorBarby, Pascal
dc.contributor.authorLacoux, Caroline
dc.contributor.authorFassy, Julien
dc.contributor.authorVial, Pablo
dc.contributor.authorVial, Cecilia
dc.contributor.authorMartínez , Jose
dc.contributor.authorOlalekan, Olusola
dc.contributor.authorInuwa, Bitrus
dc.contributor.authorShittu, Ismaila
dc.contributor.authorMeseko, Clement
dc.contributor.authorChammas, Roger
dc.contributor.authorFerreira, Carlos
dc.contributor.authorDionísio, Thiago
dc.contributor.authorGarbieri, Thais
dc.contributor.authorParisi, Aparecida
dc.contributor.authorMendes, Maria
dc.contributor.authorDe Paula, Anderson
dc.contributor.authorRomano, Camila
dc.contributor.authorBentim, Luiz
dc.contributor.authorMinoprio, Paola
dc.contributor.authorCampos, Angélica
dc.contributor.authorCunha, Marielton
dc.contributor.authorVilela, Ana
dc.contributor.authorNyirenda, Tonney
dc.contributor.authorSawasawa, Rajhab
dc.contributor.authorMuula, Adamson
dc.contributor.authorDumm, Rebekah
dc.contributor.authorHarris, Rebecca
dc.contributor.authorMitchell, Constance
dc.date.accessioned2023-05-02T16:09:29Z
dc.date.available2023-05-02T16:09:29Z
dc.date.issued2022
dc.descriptionPettit, Syril; Botten, Jason; Jerome, Keith
dc.description.abstractReverse transcription-quantitative polymerase chain reaction (RT-qPCR) is used worldwide to test and trace the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). "Extraction-less" or "direct" real time-reverse transcription polymerase chain reaction (RT-PCR) is a transparent and accessible qualitative method for SARS-CoV-2 detection from nasopharyngeal or oral pharyngeal samples with the potential to generate actionable data more quickly, at a lower cost, and with fewer experimental resources than full RT-qPCR. This study engaged 10 global testing sites, including laboratories currently experiencing testing limitations due to reagent or equipment shortages, in an international interlaboratory ring trial. Participating laboratories were provided a common protocol, common reagents, aliquots of identical pooled clinical samples, and purified nucleic acids and used their existing in-house equipment. We observed 100% concordance across laboratories in the correct identification of all positive and negative samples, with highly similar cycle threshold values. The test also performed well when applied to locally collected patient nasopharyngeal samples, provided the viral transport media did not contain charcoal or guanidine, both of which appeared to potently inhibit the RT-PCR reaction. Our results suggest that direct RT-PCR assay methods can be clearly translated across sites utilizing readily available equipment and expertise and are thus a feasible option for more efficient COVID-19 coronavirus disease testing as demanded by the continuing pandemic.
dc.description.versionVersión Publicada
dc.identifier.citationMills MG, Bruce E, Huang ML, Crothers JW, Hyrien O, Oura CAL, Blake L, Brown Jordan A, Hester S, Wehmas L, Mari B, Barby P, Lacoux C, Fassy J, Vial P, Vial C, Martinez JRW, Oladipo OO, Inuwa B, Shittu I, Meseko CA, Chammas R, Santos CF, Dionísio TJ, Garbieri TF, Parisi VA, Mendes-Correa MC, de Paula AV, Romano CM, Góes LGB, Minoprio P, Campos AC, Cunha MP, Vilela APP, Nyirenda T, Mkakosya RS, Muula AS, Dumm RE, Harris RM, Mitchell CA, Pettit S, Botten J, Jerome KR. An international, interlaboratory ring trial confirms the feasibility of an extraction-less "direct" RT-qPCR method for reliable detection of SARS-CoV-2 RNA in clinical samples. PLoS One. 2022 Jan 13;17(1):e0261853. doi: 10.1371/journal.pone.0261853
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0261853
dc.identifier.urihttps://repositorio.udd.cl/handle/11447/7432
dc.language.isoen
dc.subjectCOVID-19 / diagnosis
dc.subjectCOVID-19 / virology
dc.subjectCOVID-19 Testing / methods
dc.subjectFeasibility Studies
dc.subjectHumans
dc.subjectNasopharynx / virology
dc.subjectPandemics / prevention & control
dc.subjectRNA, Viral / genetics
dc.subjectReal-Time Polymerase Chain Reaction / methods
dc.subjectReverse Transcription / genetics
dc.subjectSARS-CoV-2 / genetics
dc.subjectSensitivity and Specificity
dc.subjectSerologic Tests / methods
dc.subjectSpecimen Handling / methods
dc.titleAn international, interlaboratory ring trial confirms the feasibility of an extraction-less "direct" RT-qPCR method for reliable detection of SARS-CoV-2 RNA in clinical samples
dc.typeArticle
dcterms.accessRightsAcceso Abierto
dcterms.sourcePloS one
dspace.entity.typePublication

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