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J Med Microbiol 54 (2005), 731-739; DOI: 10.1099/jmm.0.46058-0
© 2005 Society for General Microbiology
ISSN 0022-2615

16S rDNA PCR and denaturing gradient gel electrophoresis; a single generic test for detecting and differentiating Mycoplasma species

Laura McAuliffe1, Richard J Ellis2, Jo R Lawes1, Roger D Ayling1 and Robin AJ Nicholas1

1Mycoplasma Group, Department of Statutory and Exotic Bacterial Diseases, Veterinary Laboratories Agency, Weybridge, Surrey KT15 3NB, UK 2NERC Centre for Population Biology, Imperial College London, Silwood Park Campus, Ascot, Berkshire SL5 7PY, UK

Correspondence Laura McAuliffe l.mcauliffe{at}vla.defra.gsi.gov.uk

Received February 25, 2005
Accepted May 2, 2005

Diagnosis of Mycoplasma infection is normally based on culture and serological tests, which can be time-consuming and laborious. A number of specific PCRs have been developed but to date there has not been a single generic test capable of detecting and differentiating mycoplasmas to a species level. This report describes the development of a new diagnostic test based on PCR of the 16S rRNA gene with Mycoplasma-specific primers and separation of the PCR product according to primary sequence using denaturing gradient gel electrophoresis (DGGE). DGGE enabled the differentiation of 67 Mycoplasma species of human and veterinary origin and represents a significant improvement on current tests as diagnosis of Mycoplasma infection can be made directly from clinical samples in less than 24 h.

Abbreviations: DGGE, denaturing gradient gel electrophoresis; IGS, intergenic spacer.




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