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Peer-reviewed veterinary case report

Multiplex PCR for simultaneous detection and differentiation of sheeppox, goatpox and orf viruses from clinical samples of sheep and goats.

Journal:
Journal of virological methods
Year:
2014
Authors:
Venkatesan, G et al.
Affiliation:
Indian Veterinary Research Institute · India

Plain-English summary

Researchers have created a new test called multiplex PCR that can quickly identify three different viruses that cause skin infections in sheep and goats: sheeppox, goatpox, and orf. This test works by looking for specific parts of the viruses' genetic material and can detect very small amounts of the viruses in samples. They tested this method on 235 samples from various regions and found it to be more accurate than previous tests used in labs. This new test could help veterinarians better diagnose these infections in sheep and goats. Overall, the new test shows promise for improving the diagnosis of these viral infections.

Abstract

A multiplex polymerase chain reaction (mPCR) was developed and evaluated for detection of pox viral infections simultaneously using clinical samples from sheep and goats. Specific primers for three pox viruses of sheep and goats including sheeppox virus (SPPV), goatpox virus (GTPV) and orf virus (ORFV) were designed targeting conserved sequences of the DNA binding phosphoprotein (I3L) coding gene of Capripoxvirus (CaPV) and the DNA polymerase (E9L) gene of parapoxvirus for identification of these viruses. The mPCR assay was found to be sensitive for detecting as low as 350 pg of viral genomic DNA or 10(2) copies of standard plasmid of individual targets; and 10(3) copies of plasmid in a mixture of two or three viruses. The assay was specific for detecting one or more of the viruses in various combinations from clinical specimens. Two hundred and thirty five (n=235) clinical samples from sheep and goats received from different geographical regions of the country for diagnosis of pox infection were evaluated by developed uniplex and mPCR assays. The assay had improved diagnostic sensitivity and specificity over to in-use laboratory diagnostic methods and can be useful for clinical differential diagnosis of these infections in sheep and goats.

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Original publication: https://pubmed.ncbi.nlm.nih.gov/24134940/