Polymerase chain reaction optimization for Mycoplasma gallisepticum and M. synoviae diagnosis.
Mycoplasma gallisepticum- or M. synoviae-challenged chickens were monitored with serological assays (serum plate agglutination, hemagglutination inhibition, and enzyme-linked immunosorbent assay) and polymerase chain reaction (PCR). The tracheal swabs from M. gallisepticum-challenged chickens received three different treatments (phosphate-buffered saline [PBS], Frey's broth, or 10 mM Tris-HCl/250 mM ethylenediaminetetraacetic acid/ 2.5% sodium dodecyl sulfate [STE]) prior to DNA purification. A nonphenolic method for DNA extraction was utilized. The best PCR results were obtained with PBS swab treatment. The nonphenolic method for DNA extraction was compared with a phenolic method in an experiment with tracheal swabs from M. synoviae-challenged chickens and commercial flocks. Both methods gave comparable results.
Silveira
R M
RM
Departamento de Bioquímica, UFRGS Porto Alegre, Rio Grande do Sol, Brazil.
Fiorentin
L
L
Marques
E K
EK
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D004269:DNA, Bacterial / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
D004269:DNA, Bacterial