Rectovaginal group B Streptococcus (GBS) colonisation in pregnant individuals at the time of labour is a major risk factor for invasive GBS disease by age 7days (early-onset disease). We aimed to investigate the prevalence of rectovaginal GBS colonisation at the time of labour among pregnant women and vertical transmission to their newborns across selected low-income and middle-income African and south Asian countries. This prospective, observational study was undertaken at 11maternity and obstetric care facilities based in Ethiopia, Kenya, Mozambique, Nigeria, Mali, South Africa, Bangladesh, India, and Bhutan. HIV-negative pregnant women aged 18-45years who were in the early stages of labour and at least 37weeks' gestation were eligible for inclusion. Lower vaginal and rectal swabs and urine were collected from the women, and swabs of the umbilicus, outer ear, axillary fold, rectum, and throat were obtained from their newborns, for GBS culture. Standardised sampling and culture using direct plating and selective media broth for detection of GBS colonisation was undertaken at the sites. Serotyping of GBS isolates was done in South Africa. The primary outcome was the prevalence of rectovaginal GBS among pregnant women, analysed in participants with available data. This study is registered with the South African National Clinical Trials Register, number DOH-27-0418-4989. 6922pregnant women were enrolled from Jan 10, 2016, to Dec 11, 2018, of whom 6514 (94·1%; 759-892per country) were included in the analysis; data from Bhutan were not included in the study due to issues with specimen collection and processing. Overall, the prevalence of maternal GBS colonisation was 24·1% (95% CI 23·1-25·2; 1572of 6514); it was highest in Mali (41·1% [37·7-44·6]; 314of 764) and lowest in Ethiopia (11·6% [9·5-14·1]; 88of 759). Theoverall rate of vertical transmission of GBS from women with rectovaginal GBS colonisation was 72·3% (70·0-74·4; 1132of 1566); it was highest in Mozambique (79·2% [73·3-84·2]; 168of 212) and lowest in Bangladesh (55·8%, 47·5-63·8; 77of 138). The five most common GBS colonising serotypes were Ia (37·3% [34·9-39·7]; 586of 1572), V(28·5% [26·3-30·8]; 448of 1572), III (25·1% [23·0-27·3]; 394of 1572), II (9·2% [7·8-10·7]; 144of 1572), and Ib(6·5% [5·4-7·8]; 102of 1572). There was geographical variability in serotype proportion distribution; serotype VIIwas the third most common serotype in India (8·6% [5·3-13·7]; 15of 174) and serotype VI was mainly identified in Bangladesh (5·8% [3·0-11·0]; eight of 138) and India (5·7% [3·2-10·3]; ten of 174). Our study reported a high prevalence of GBS colonisation in most settings, with some geographical variability even within African countries. Our findings suggest that serotypes not included in current multivalent capsular-polysaccharide GBS vaccines prevail in some regions, so vaccine efficacy and post-licensure effectiveness studies should assess the effect of vaccination on maternal GBS colonisation given the potential for replacement by non-vaccine serotypes. Bill & Melinda Gates Foundation.