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Query: UMLS:C0017160 (gastroenteritis)
 11,398 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

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.
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PMID:Molecular-based identification and typing of Campylobacter jejuni and C. coli. 1515 16

Campylobacter species are the leading agents of bacterial gastroenteritis worldwide. C. jejuni and C. coli together are responsible for more than 95% of all cases of Campylobacter-induced diarrheal disease in the United States. Detection of campylobacteria in clinical samples by conventional culture is problematic and slow due to their complex taxonomy, fastidious growth requirements, and biochemical inertness. The current study describes a rapid, sensitive, and specific real-time polymerase chain reaction (PCR) assay capable of detecting and differentiating C. jejuni (hippuricase gene, hipO) and C. coli (serine hydroxymethyltransferase gene, glyA) in a single reaction, directly from clinical isolates and human feces. The analytical specificity of the assay was demonstrated with a diverse range of Campylobacter species, related organisms, and other diarrhea-inducing bacterial pathogens. The analytical sensitivity of the multiplexed, PCR assay was 10 genome copies for both C. jejuni and C. coli. Following a rapid DNA extraction method (QIAGEN QIAamp DNA stool Mini Kit), the multiplexed PCR identified C. jejuni or C. coli in 100% of fecal samples containing 10(3) colony-forming units (CFU) per gram of feces. This assay represents the first real-time PCR method capable of detecting and differentiating C. jejuni and C. coli in a single reaction.
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PMID:A real-time multiplexed PCR assay for rapid detection and differentiation of Campylobacter jejuni and Campylobacter coli. 1527 89

Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.
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PMID:Differentiation of Campylobacter jejuni and Campylobacter coli Using Multiplex-PCR and High Resolution Melt Curve Analysis. 2639 42