The subject of the paper focuses on the effect of weave architecture on microbial barrier properties of woven fabrics or more precisely on identifying crucial elements of weave architecture that dominantly influence bacteria penetration in dry condition. For that purpose, 12 samples of cotton fabrics were woven and examined. In their structure, all samples had the same yarns (36 tex) in warp and weft, same densities of warp (24 yarns/cm), two weft densities (24 and 20 yarns/cm) and six different basic weave structures. Microbial barrier permeability was determined according to a previously developed test method in cooperation with University Hospital Center Zagreb. Bacterial endospores of apathogenic species of the genus Bacillus: Geobacillus stearothermophilus and Bacillus atrophaeus were used. The effect of weave pattern on microbial barrier properties was significant. Weave patterns, decisively determined the number of influencing pores and its sizes in woven fabrics, as well as the yarn floating which jointly almost perfectly correlated with bacteria penetration through the woven fabric. Multiple linear regression of pore numbers and floating threads produced equations which correspond in 99% to the measuring results for densities 24/24 and 24/20, and more than 98% considering both densities of the set. Among compared weave patterns, satin weave had significantly lower permeability of microorganisms (six–seven times) than basket weave (the highest), for both densities.