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Rapid identification of Campylobacter jejuni strains by polymerase chain reaction & their restriction fragment length polymorphism analysis. 1997

M Mahendru, and K N Prasad, and T N Dhole, and A Ayyagari
Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow.

A polymerase chain reaction (PCR) technique was developed for specific identification of C. jejuni. A primer pair of a conserved region of flagellin A (fla A) gene identified all 15 strains of C. jejuni isolated from human faeces. None of the control strains like Helicobacter pylori, Vibrio cholerae Escherichia coli and Salmonella typhimurium except C. coli exhibited any amplified product by PCR. A predicted 450 bp could also be amplified from 4 chicken caecal contents positive for C. jejuni-C coli by culture. The caecal contents remained positive for C. jejuni-C. coli by PCR after preservation at 4 degrees C for one week when no viable organism could be detected. Restriction fragment length polymorphism analysis (RFLP) of fla A amplified product by using Bgl II enzyme classified 15 strains into 5 types. Three C. jejuni strains isolated from the same patient over a period of 3 wk showed the same RFLP pattern. The present study indicates that PCR is specific for C. jejuni-C. coli and it has the potential for rapid diagnosis of infection. RFLP can be a good epidemiological marker for C. jejuni infection.

UI MeSH Term Description Entries
D012150 Polymorphism, Restriction Fragment Length Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment. RFLP,Restriction Fragment Length Polymorphism,RFLPs,Restriction Fragment Length Polymorphisms
D005243 Feces Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D016123 Campylobacter jejuni A species of bacteria that resemble small tightly coiled spirals. Its organisms are known to cause abortion in sheep and fever and enteritis in man and may be associated with enteric diseases of calves, lambs, and other animals. Campylobacter fetus subsp. jejuni,Vibrio hepaticus,Vibrio jejuni
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain

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