Molecular-based identification and typing of Campylobacter jejuni and C. coli.
Thermophilic Campylobacter spp., mainly Campylobacter jejuni and to a lesser extent C. coli are recognized as the most common bacteriological causes of gastroenteritis in humans. As enteric infection with Campylobacter organisms cannot be distinguished from that caused by other enteric pathogens, a definitive diagnosis can only be made by isolating or detecting the organism from the feces. The epidemiology of Campylobacter enteritis has been complicated by the ubiquitous nature of the organism (commonly found as a commensal in the intestines of domestic animals, in milk, and in water). Furthermore, identification is carried out only to genus level by most clinical laboratories. Because of the biochemical similarity known to exist between C. jejuni and C. coli, the hippurate hydrolysis test is often used as the only phenotypic test capable of differentiating the two species. This test, however, has some acknowledged technical limitations and is dependent on inoculum size; results can be difficult to interpret accurately. Furthermore, almost all C. jejuni isolates possess the hippuricase gene, fewer C. jejuni isolates express the hippuricase gene. For this reason, certain polymerase chain reaction (PCR)-based species identification methods, for both C. jejuni and C. coli, and for the other thermophilic species, provide more reliable identification; they also help to highlight mixed species cultures, should they occur. However, even with these methods, false negatives or nonspecifically amplified product(s) can occur in a minority of isolates tested owing to genomic anomalies. Thus a second molecular identification method may be required in these circumstances. Gonzalez et al. developed a species-specific PCR assay for the identification of C. jejuni and C. coli based on the ceuE gene, which is involved in siderophore transport. Using this method two primer sets are employed in separate PCR amplification reactions. Another method, developed by Eyers et al., performs PCR amplification of 23S rRNA gene fragments, based on regions specific for C. jejuni, C. coli, C. lari, and C. upsaliensis. In addition, Hani and Chan developed a PCR assay that detected and amplified the hippuricase gene. This molecular approach may offer a more reliable means of identifying C. jejuni strains compared with the phenotypic hippurate hydrolysis test alone.
Lucey
Brigid
B
Molecular Diagnostic Unit, Cork Institute of Technology, Bishoptown, Cork, Ireland.
O'Halloran
Fiona
F
Fanning
Séamus
S
D015373:Bacterial Typing Techniques / Q000379:methods*; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004586:Electrophoresis / Q000379:methods; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods
D017931:DNA Primers; D004269:DNA, Bacterial