The chicken major histocompatibility complex B (MHC-B) – a cluster of genes associated with natural disease resistance or susceptibility – has been investigated in experimental and inbred lines by serological typing. However, that method showed some weaknesses for its use on out breeds. This study aims to explore the genetic diversity of the MHC-B of Cameroon indigenous chicken through molecular typing with LEI0258 and MCW0371 microsatellite markers. MHC-B haplotypes of 290 chickens from four agro-ecological zones were identified and compared with published haplotypes. Alleles were analysed for genetic diversity and relationship among Cameroon chicken populations. Hypothetically new LEI0258 alleles and haplotypes were detected. Overall, polymorphism parameters were relatively high in the Cameroon western highlands. The analysis of molecular variance revealed great variability (80.00 percent) between individuals than among and within ecotypes. The inbreeding coefficients of overall populations (F
IT), among population (F
ST) and within population (F
IS) were 0.26, 0.04 and 0.22, respectively, and all were highly significant (P < 0.001). A UPGMA tree based on Nei's D
A genetic distances showed a clear distinction between Cameroon and out-groups and a structuring of within-country populations into three clusters. There is a great genetic diversity of the MHC-B in Cameroon native chicken and also a need of sequencing of the identified alleles for an accurate identification prior to their assessment for natural disease resistance and responsiveness to vaccination.