Detection of 12 respiratory viruses by duplex real time PCR assays in respiratory samples

Molecular and Cellular Probes
Rosaria ArviaAlberta Azzi

Abstract

Different viruses can be responsible for similar clinical manifestations of respiratory infections. Thus, the etiological diagnosis of respiratory viral diseases requires the detection of a large number of viruses. In this study, 6 duplex real-time PCR assays, using EvaGreen intercalating dye, were developed to detect 12 major viruses responsible for respiratory diseases: influenza A and B viruses, enteroviruses (including enterovirus spp, and rhinovirus spp), respiratory syncytial virus, human metapneumovirus, coronaviruses group I (of which CoV 229E and CoV NL63 are part) and II (including CoV OC43 and CoV HKU1), parainfluenza viruses type 1, 2, 3 and 4, human adenoviruses and human bocaviruses. The 2 target viruses of each duplex reaction were distinguishable by the melting temperatures of their amplicons. The 6 duplex real time PCR assays were applied for diagnostic purpose on 202 respiratory samples from 157 patients. One hundred fifty-seven samples were throat swabs and 45 were bronchoalveolar lavages. The results of the duplex PCR assays were confirmed by comparison with a commercial, validated, assay; in addition, the positive results were confirmed by sequencing. The analytical sensitivity of the duplex PCR assays vari...Continue Reading

References

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Citations

May 4, 2016·Medical Microbiology and Immunology·Manuela BonizzoliAdriano Peris
Dec 28, 2017·The Journal of Infectious Diseases·Marie L Landry, Ellen F Foxman
Aug 9, 2020·Medical Hypotheses·Bruno Sposato, Marco Scalese

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Methods Mentioned

BETA
PCR
bronchoalveolar lavage

Software Mentioned

ClustalW
BioEdit

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