Colonization factors (CFs) mediate attachment of enterotoxigenic Escherichia coli (ETEC) to the intestinal mucosa and induce protective immunity against ETEC diarrhoea. We designed CF-specific polymerase chain reaction (PCR) primers, and developed a simple PCR-based genotypic CF identification method. ETEC strains (n=17) isolated from patients with diarrhoea in Thailand were examined for genotypical identification of CFs of ETEC strains. Coli surface antigen 6 (CS6) was the most common CF (29%), followed by CS13 (12%), colonization factor antigen I (CFA/I), CS2 and CS3, and CS17/CS19 (6% each), while 41% of the strains were negative. This simple PCR method for the detection of CF genes is useful for surveillance of ETEC infections in diagnostic laboratories.