Detection and molecular typing of Campylobacter jejuni in fecal samples by polymerase chain reaction.
In order to determine whether polymerase chain reaction (PCR) could be used to detect Campylobacter jejuni directly in stool samples, DNA from 66 frozen culture positive and negative fecal samples was purified by column chromatography. The flaA gene was amplified using primers directed against the conserved 5' and 3' regions and produced a 1.7 kb amplicon. Fifteen of 18 (83%) C. jejuni culture positive samples were detected by agarose gel electrophoresis and ethidium bromide staining. The test was negative in one sample containing C. coli. Twelve samples containing other enteric pathogens were negative as were 34 of 35 culture negative samples. Flagellin gene typing (see reference 14) of the flaA gene product from two stool samples in which the patients' stool isolate was also available showed the identical flagellin gene types suggesting that molecular typing of Campylobacter could potentially be performed on stool samples without the need for culture.
Waegel
A
A
Department of Biology, Neumann College, Aston, Pennsylvania, USA.
Nachamkin
I
I
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D002169:Campylobacter Infections / Q000175:diagnosis* / Q000382:microbiology; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D003937:Diagnosis, Differential; D005243:Feces / Q000382:microbiology; D005408:Flagellin / Q000235:genetics*; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies
D017931:DNA Primers; D005408:Flagellin