MOLECULAR DISCRIMINATION BETWEEN CAMPYLOBACTER-JEJUNI, CAMPYLOBACTER-COLI, CAMPYLOBACTER-LARI AND CAMPYLOBACTER-UPSALIENSIS BY POLYMERASE CHAIN-REACTION BASED ON A NOVEL PUTATIVE GTPASE GENE

Polymerase chain reaction (PCR) mediated DNA fingerprinting has result ed in the identification of a novel Campylobacter jejuni gene, encodin g a GTPase protein. The gene, consisting of 383 amino acids contained semi-conserved CTP-binding sites (designated G-1 to G-4), that are cha racteristic for members of the CTPase protein superfamily. Remarkably, this gene from C. jejuni appears to encode a member of a novel family of GTP-binding proteins, containing two separate putative CTP-binding domains, each comprising a series of semi-conserved GTP-binding motif s. Spacing between these motifs is highly conserved. Based on this nov el gene, a general PCR strategy for the identification of C. jejuni, C . coil, C. lari and C. upsaliensis was developed. PCR primers were ded uced from GTP-binding motifs G-1 and G-3 of the first GTP-biriding dom ain. These GTP-binding sites flank a variable region of precisely 117 bp in the four Campylobacter spp. that allowed the development of spec ies-specific probes. This PCR-hybridization assay offers a novel tool for rapid molecular detection and specific identification of the therm ophilic Campylobacter spp. (C) 1997 Academic Press Limited.