Detection of pyrazinamidase activity for differentiation of Campylobacter, Arcobacter, and Helicobacter spp. by using a high-performance liquid chromatography method

A high-performance liquid chromatography method was investigated for the detection pyrazinamidase activity by Campylobacter, Arcobacter, and Helicobacter spp. Pyrazine carboxilic acid, one of the end products of pyrazinamide hydrolysis by microorganisms, was detected by using a high-performance liquid chromatography (HPLC). A loopful of organism colonies was emulsified in 0.5 ml of a 0.5% pyrazinamide solution. The suspens on was incubated in a 37 degrees C water bath for 18-20 hr. After centrifugation, the supernatant was analyzed by HPLC. This HPLC method does not require microaerobic incubation and was easy to interpret for strains with weak enzymatic activity. By this method, we tested 111 clinical isolates, type and reference strains of Campylobacter spp., Arcobacter spp., and Helicobacter spp. , C. jejuni, C. jejuni subsp. doylei, C. coli, C. upsaliensis, C. lari, C. lari (urease+), C. helveticus, C. hyolei, C. sputorum subsp. fecalis, C. gracilis, C. concisus, C. curvus were positive for pyrazinamidase. C. fetus subsp. fetus, C. hyointestinalis, C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, C. mucosalis, A. butzleri, A. skirrowii, A. cryaerophilus, H. pylori, H. cinaedi, H. fennelliae, H. mustelae, H. felis, H. muridarum, H. canis, H. nemestrinae, H. pamentensis, H. pullourum were negative.