Development of a sensitive nested PCR method for the specific detection of Mycoplasma mycoides subsp. mycoides SC

Molecular and Cellular Probes
R MiserezJ Frey

Abstract

A specific and sensitive test for the detection of Mycoplasma mycoides subsp. mycoides small colony type (SC), the aetiological agent of contagious bovine pleuropneumonia (CBPP) was developed using two nested PCR reactions. The PCR reactions are based on the nucleotide sequence of lipoprotein P72 of M. mycoides subsp. mycoides SC. The two specific oligonucleotide primer pairs were chosen to match those sequence segments of the P72 gene which differ most from the gene of the closely related lipoprotein P67 of Mycoplasma sp. bovine group 7 (strain PG50). The nested PCR reacted with all of the 34 different strains of M. mycoides subsp. mycoides SC analysed, and gave no amplification product with any of the closely related mycoplasmas tested, showing its high specificity. In bronchial lavage fluid experimentally contaminated with M. mycoides subsp. mycoides SC, the assay was able to detect as few as two viable cells per ml using a simple lysis procedure prior to the amplification step. With clinical samples, the sensitivity of the nested PCR was about 10(4)-10(5) higher than that of single PCR amplifications performed under the same conditions. The assay was also successfully used to detect M. mycoides subsp. mycoides SC in bronchi...Continue Reading

References

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Citations

May 31, 2011·Preventive Veterinary Medicine·Florence TardyFrançois Poumarat
Sep 6, 2000·Molecular and Cellular Probes·S LorenzonF Thiaucourt
Apr 19, 2006·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Katie MilesRobin A J Nicholas
Jul 7, 2000·Clinical and Diagnostic Laboratory Immunology·E M AbdoJ Frey
Oct 4, 2007·International Journal of Systematic and Evolutionary Microbiology·Lucía Manso-SilvánFrançois Thiaucourt
Feb 17, 1998·Applied and Environmental Microbiology·K D StärkJ Frey
Mar 18, 2003·The Veterinary Record·B CetinkayaF Thiaucourt
May 23, 1998·Journal of Clinical Microbiology·D F TalkingtonJ S Jensen

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