Development of fast and sensitive protocols for the detection of viral pathogens using a small portable convection PCR platform

One of the most crucial steps for preventing viral pandemics is the early detection of the causative virus on site. Various molecular and immunological approaches have been developed for virus detection. In this study, we investigated the utility of the recently introduced convection polymerase chai...

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Bibliographic Details
Published in:Molecular biology reports 2019-10, Vol.46 (5), p.5073-5077
Main Authors: Lee, Myoung Hui, Song, Kyung-Young, Hwang, Hyun Jin, Kim, Jeong Hee, Hwang, Inhwan
Format: Article
Language:English
Subjects:
Animal Anatomy
Animal Biochemistry
Animals
Avian flu
Biomedical and Life Sciences
Birds - genetics
Convection
DNA Primers - genetics
Foot & mouth disease
Foot-and-Mouth Disease Virus - genetics
Hemagglutinins
Histology
Influenza A virus - genetics
Influenza in Birds - diagnosis
Life Sciences
Morphology
Original Article
Pandemics
Polymerase chain reaction
Polymerase Chain Reaction - instrumentation
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Virus Diseases - diagnosis
Virus Diseases - genetics
Viruses
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Summary:One of the most crucial steps for preventing viral pandemics is the early detection of the causative virus on site. Various molecular and immunological approaches have been developed for virus detection. In this study, we investigated the utility of the recently introduced convection polymerase chain reaction (cPCR) platform for the rapid and sensitive detection of various animal viruses in the field, including the foot-and-mouth disease virus (FMDV) and avian influenza viruses (AIVs). Primer sets were designed to simultaneously detect two highly conserved regions of the FMDV, including the 5′ untranslated region (5′-UTR) and 3D gene, and to specifically amplify the NP and hemagglutinin ( HA ) genes of H5 and H9 subtypes of AIVs. The portable cPCR system was able to amplify from as low as 1 to 10 copies of viral cDNAs in the singleplex mode and 10 to 100 copies of viral cDNAs in the duplex mode within 21 min. Thus, our data suggest that the cPCR protocols developed in this study are highly sensitive and enable quick detection of animal viruses in biological samples.
ISSN:0301-4851
1573-4978
DOI:10.1007/s11033-019-04961-x