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Rinderpest virus lineage differentiation using RT-PCR and SNAP-ELISA

Version 2 2024-06-04, 06:34
Version 1 2017-05-17, 14:31
journal contribution
posted on 2024-06-04, 06:34 authored by Maria ForsythMaria Forsyth, S Parida, Soren AlexandersenSoren Alexandersen, GJ Belsham, T Barrett
An RT-PCR/ELISA system has been developed that detects and differentiates Rinderpest virus (RPV) from the other closely related morbillivirus of ruminants, Peste des petits Ruminants virus (PPRV). In addition, using lineage specific probes, it is possible to determine whether the virus sample is wild-type or vaccine, and the likely origin of the outbreak if it is wild-type. It involves carrying out a RT-PCR with one digoxygenin (Dig)-labelled primer followed by a hybridisation step with a virus-specific, biotin-labelled, probe. The hybridisation step is carried out in an ELISA format on a streptavidin-coated plate. The DIG-labelled products are detected using a specific anti-DIG monoclonal antibody and an anti-mouse horseradish peroxidase conjugate. The hybridisation step replaces nucleotide sequencing or nested PCR for confirmation of the identity of DNA product. The assay is fast and easy to carry out and can give semi-quantitative estimates of the virus content of samples.

History

Journal

Journal of virological methods

Volume

107

Pagination

29-36

Location

Netherlands

ISSN

0166-0934

Language

eng

Publication classification

CN.1 Other journal article

Issue

1

Publisher

Elsevier