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 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.
Larrasa
José
J
Departamento de Microbiología, Laboratorios Larrasa S.L., Corredera Hernando de Soto 13-A, Jerez de los Caballeros, 06380 Badajoz, Spain.
García-Sánchez
Alfredo
A
Ambrose
Nicholas C
NC
Parra
Alberto
A
Alonso
Juan M
JM
Rey
Joaquín M
JM
Hermoso-de-Mendoza
Miguel
M
Hermoso-de-Mendoza
Javier
J
D000192:Actinomycetales / Q000145:classification* / Q000235:genetics*; D000193:Actinomycetales Infections / Q000382:microbiology* / Q000662:veterinary; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology; D003670:Deer; D015252:Deoxyribonucleases, Type II Site-Specific; D003872:Dermatitis / Q000382:microbiology* / Q000662:veterinary; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods* / Q000592:standards; D005798:Genes, Bacterial; D006736:Horses; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D015183:Restriction Mapping; D012756:Sheep; D012757:Sheep Diseases / Q000382:microbiology
C066157:endodeoxyribonuclease Sse83871; D015252:Deoxyribonucleases, Type II Site-Specific