Purpose: to indentify the prevalence and risk factors of maternal colonization by group B streptococcus (GBS) in pregnant women with premature labor (PL) and/or premature membrane rupture (PMR). METHODS: two anal and two vaginal swabs were collected from 203 pregnant women with diagnosis of PL or PMR assisted at the practice along one year. Pregnant women with imminent labor at admission were excluded. One swab of each source was placed in a transfer milieu and sent for culture in blood-agar plates; the two remaining swabs were incubated for 24 hours in Todd-Hewitt milieu for further sowing in blood-agar plates. Risk factors were analyzed by the chi-square test, Student's t-test (p-value set at 0.05 and 95% confidence interval) and logistic regression. The following variables were analyzed: age, race, parity and mother schooling; culture results by source and type of culture; admission diagnosis; gestational age at admission; asymptomatic bacteriuria; gestational age at delivery; type of delivery; neonatal GBS colonization rate and immediate neonatal condition. RESULTS: prevalence of maternal GBS colonization was 27.6% (56 cases). The colonization rates according to gestational complications were 30% for PMR, 25.2% for PL and 17.8% for PL + PMR. Univariate analysis has shown that the variables Caucasian race, low level of schooling and bacteriuria were associated with higher colonization rates. Multivariate analysis showed that the presence of urinary infection was the only variable associated with maternal colonization. The GBS detection rate was significantly higher with the use of a selective milieu and collection from both anal and vaginal sources. The neonatal colonization rate was 3.1%. Two cases of early sepsis by GBS occurred in the sample, with prevalence of 10.8 cases per one thousand live births and 50% mortality rate. Conclusion: the studied sample showed high maternal colonization rates by Streptococcus agalactiae. To increase GBS detection rate, it is necessary to use a selective culture milieu and to combine anal-rectal and vaginal cultures. 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