Evaluation of randomly amplified polymorphic DNA and pulsed field gel electrophoresis techniques for molecular typing of Dermatophilus congolensis

This study aimed to evaluate molecular typing methods useful for standardization of strains in experimental work on dermatophilosis. Fifty Dermatophilus congolensis isolates, collected from sheep, cattle, horse and a deer, were analyzed by randomly amplified polymorphic DNA (RAPD) method using twent...

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Bibliographic Details
Published in:FEMS microbiology letters 2004-11, Vol.240 (1), p.87-97
Main Authors: Larrasa, J, Garcia-Sanchez, A, Ambrose, N.C, Parra, A, Alonso, J.M, Rey, J.M, Hermoso-de-Mendoza, M, Hermoso-de-Mendoza, J
Format: Article
Language:English
Subjects:
Actinomycetales - classification
Actinomycetales - genetics
Actinomycetales Infections - microbiology
Actinomycetales Infections - veterinary
Amplification
animal pathogenic bacteria
Animals
Bacteriology
Biological and medical sciences
Cattle
Cattle Diseases - microbiology
Deer
Deoxyribonucleases, Type II Site-Specific
Deoxyribonucleic acid
dermatitis
Dermatitis - microbiology
Dermatitis - veterinary
Dermatophilus
Dermatophilus congolensis
DNA
Electrophoresis
Electrophoresis, Gel, Pulsed-Field - methods
Electrophoresis, Gel, Pulsed-Field - standards
Epidemiology
exudative dermatitis
Fundamental and applied biological sciences. Psychology
Gel electrophoresis
Genes, Bacterial
Genetics
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Horses
Microbiology
Molecular typing
Mycology
PFGE
provenance
pulsed-field gel electrophoresis
random amplified polymorphic DNA technique
Random Amplified Polymorphic DNA Technique - methods
Random Amplified Polymorphic DNA Technique - standards
RAPD
Reproducibility
Reproducibility of Results
Restriction Mapping
Sheep
Sheep Diseases - microbiology
Standardization
Streptotrichosis
Typing
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Summary:This study aimed to evaluate molecular typing methods useful for standardization of strains in experimental work on dermatophilosis. Fifty Dermatophilus congolensis isolates, collected from sheep, cattle, horse and a deer, were analyzed by randomly amplified polymorphic DNA (RAPD) method using twenty-one different primers, and the results were compared with those obtained by typing with a pulsed field gel electrophoresis (PFGE) method using the restriction digest enzyme Sse8387I. The typeability, reproducibility and discriminatory power of RAPD and Sse8387I-PFGE typing were calculated. Both typing methods were highly reproducible. Of the two techniques, Sse8387I-PFGE was the least discriminating (Dice Index (DI), 0.663) and could not distinguish between epidemiologically related isolates, whereas RAPD showed an excellent discriminatory power (DI, 0.7694-0.9722). Overall, the degree of correlation between RAPD and PFGE typing was significantly high (r, 0.8822). We conclude that the DNA profiles generated by either RAPD or PFGE can be used to differentiate epidemiologically unrelated isolates. The results of this study strongly suggest that at least two independent primers are used for RAPD typing in order to improve its discriminatory power, and that PFGE is used for confirmation of RAPD results.
ISSN:0378-1097
1574-6968
DOI:10.1016/j.femsle.2004.09.016