Differentiation of mycoplasma gallisepticum strains using molecular methods.
Increasing use of Mycoplasma gallisepticum (MG) live vaccines has led to a need for the differentiation of MG strains. The MG strains MK-7, MS-16, S6, FS-9 and R strains and the MG live vaccine strain F were compared by random amplification of polymorphic DNA (RAPD) in this study. Using RAPD, different patterns were found among the MG strains. In addition to this, we examined the differentiating potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) primers targeted at the crmA, crmB, crmC, gapA, mgc2 and pvpA genes encoding cytadherence-related surface proteins. These proteins may take part in the pathogenesis of MG-induced disease. Differentiation of strain F is based on the identification of restriction enzyme sites in the PCR amplicons. Using HphI enzyme, crmC PCR amplicons produced different RFLP patterns. Digestion of amplicons of gapA-specific PCR with MboI enzyme also produced distinct patterns. Differences were observed among strains R and F by digestion of mgc2 PCR amplicons with HaelIl and VspI enzymes and digestion of pvpA PCR amplicons with AccI, PvulI and ScrFI endonucleases. This method can be used for the rapid differentiation of vaccine strain from wild strains. Differentiation of MG strains is a great advantage for diagnosticians or practitioners and it is useful for epidemiological studies.
Biró
Judit
J
Veterinary Medical Research Institute of the Hungarian Academy of Sciences, H-1581 Budapest, P.O. Box 18, Hungary. bjudit@vmri.hu
Erdei
Noémi
N
Székely
Ibolya
I
Stipkovits
L
L
D000818:Animals; D001428:Bacterial Vaccines; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D009175:Mycoplasma Infections / Q000517:prevention & control / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011200:Poultry; D011201:Poultry Diseases / Q000382:microbiology / Q000517:prevention & control*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity
D001428:Bacterial Vaccines; D017931:DNA Primers; D004269:DNA, Bacterial