Use of the polymerase chain reaction to detect mycoplasma DNA in cattle with contagious bovine pleuropneumonia
Bashiruddin, J.B.; Nicholas, R.A.; Santini, F.G.; Ready, R.A.; Woodward, M.J.; Taylor, T.K.
Veterinary Record 134(10): 240-241
1994
ISSN/ISBN: 0042-4900 PMID: 8197684 Document Number: 432693
DNA was extracted from homogenised samples of a bovine lung from which Mycoplasma mycoides subspecies mycoides small colony (SC) had been isolated and from lung samples from a cow from which M. bovis had been isolated and from an animal with no known disease and also from cultures of 5 recent field isolates of M. mycoides subspecies Mycoides SC. The polymerase chain reaction (PCR) was carried out on the DNA extracts using primers which had been shown to amplify a 0.5 kb region of the genome of the M. mycoides group. The PCR products from the DNA extracted from the lung of the cow with contagious bovine pleuropneumonia and from the Afade reference strain of M. mycoides subspecies mycoides SC gave identical bands when analysed by gel electrophoresis. The digestion patterns of the PCR products of the Afade reference strain and the lung extracts produced by using the restriction enzyme Asn1 were identical and clearly distinguishable from the F30 reference strain of M. mycoides subspecies mycoides large colony (LC) form.This latter strain had an identical pattern to M. mycoides subspecies capri. The PCR products from the 5 field isolates gave the same restriction patterns as the Afade reference strain. The DNA from the uninfected control and the animal injected with M. bovis gave no bands. The results showed that the PCR can identify M. mycoides subspecies mycoides SC in bacterial isolates or clinical material within 2 days of extraction and confirmed the specificity of the test.