A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test
Joint Authors
Lamoral, Sophie
Suin, Vanessa
Francart, Aurélie
Van Gucht, Steven
Nazé, Florence
Kalai, Michael
De Craeye, Stéphane
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-04-15
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR), based on a nested PCR strategy, was validated for the detection of different lyssavirus species.
Primers with 17 to 30% of degenerate bases were used in both consecutive steps.
The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV.
In silico sequence alignment showed a functional match with the remaining lyssavirus species.
The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test.
The limit of detection was ≤1 50% tissue culture infectious dose.
The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses.
The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation.
The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration.
Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible.
American Psychological Association (APA)
Suin, Vanessa& Nazé, Florence& Francart, Aurélie& Lamoral, Sophie& De Craeye, Stéphane& Kalai, Michael…[et al.]. 2014. A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test. BioMed Research International،Vol. 2014, no. 2014, pp.1-12.
https://search.emarefa.net/detail/BIM-457838
Modern Language Association (MLA)
Suin, Vanessa…[et al.]. A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test. BioMed Research International No. 2014 (2014), pp.1-12.
https://search.emarefa.net/detail/BIM-457838
American Medical Association (AMA)
Suin, Vanessa& Nazé, Florence& Francart, Aurélie& Lamoral, Sophie& De Craeye, Stéphane& Kalai, Michael…[et al.]. A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test. BioMed Research International. 2014. Vol. 2014, no. 2014, pp.1-12.
https://search.emarefa.net/detail/BIM-457838
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-457838