MOLECULAR TYPING USING RAPD-PCR IN GROUP B STREPTOCOCCAL ISOLATED FROM NEWBORNS IN MISIONES

LACZESKI, ME; NOVOSAK, MG; BOBADILLA, FJ; ALVARENGA, AE; VERGARA, MI

Mar del Plata

Congreso; SAIB - 51 Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2015

Streptococcus agalactiae, Group B Streptococcus (GBS) is considered to be the major cause of neonatal sepsis andmeningitis of bacterial origin. The Random Amplified Polymorphic DNA analysis (RAPD) is an accessible andsensitive method based on the use of arbitrary primers to amplify polymorphic segments of DNA and a powerfulmethod to differentiate GBS strains. The aim of this study was to determine if there clonal relationship between the strains of group GBS isolated from newborns in the province of Misiones, Argentina. For this purpose eleven strainsof GBS invasive isolated from blood cultures and cerebrospinal fluid (CSF) were studied by RAPD using threeprimers, designated OPS11, OPB17, and OPB18. The PCR mixture consisted of buffer (10 mM Tris-HCl [pH 8.3],50 mM KCl; 2.5 mM MgCl2), 100 mM each of the four deoxynucleoside triphosphates (INBIO, Argentina), 0.4 mMprimer and 2.5 U of Taq DNA polymerase (INBIO, Argentina) in a total volume of 25 ml. Eleven different bandpatterns were obtained by RAPD-PCR with significant bands ranging from 100-5000 bp. In conclusion, the RAPDPCRassay showed that each of the isolated clones belonged to a different RAPD genotype, revealing that neonatalinvasive GBS infections were not epidemiologically related. The genetic diversity studies provide insight into theregional epidemiology of this disease and adapt prevention strategies.