Development of a novel digital RT-PCR method for detection of human sapovirus in different matrices
Autor: | Miguel F. Varela, Enrique Rivadulla, Ricardo Santos, Silvia Monteiro, Jesús L. Romalde |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Genotype biology Reverse Transcriptase Polymerase Chain Reaction Absolute quantification 030106 microbiology Reproducibility of Results Sapovirus Computational biology biology.organism_classification Sensitivity and Specificity Gastroenteritis 03 medical and health sciences 030104 developmental biology Real-time polymerase chain reaction Environmental water Virology biology.protein Humans RNA Viral Control material Polymerase Caliciviridae Infections |
Zdroj: | Journal of Virological Methods. 254:21-24 |
ISSN: | 0166-0934 |
Popis: | Summary A new nanofluidic digital RT-PCR method was developed for sapovirus (SaV) using control material obtained according to standards for enteric viruses. Primers employed amplify a fragment of 112 bp of the polymerase capsid junction, allowing the detection of human genogroups I, II and IV. Analytical validation was performed in clinical, shellfish and environmental water samples. This novel protocol rendered great effectiveness and repetitiveness, as well as higher sensitivity than real time RT-PCR assay, with differences in quantification ranging from 0.1 to 2.6 log-units. The method described here can constitute a promising tool for standardizing SaV quantification. |
Databáze: | OpenAIRE |
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