Development of a PCR assay combined with a short enrichment culture for detection of Campylobacter jejuni in estuarine surface waters.
Two extraction procedures were examined, and it was found that DNA recovered from Campylobacter jejuni lysed by the cetyltrimethylammonium bromide (CTAB) method was more suitable for use as a PCR template than DNA released by the boiling method. The region targeted for PCR amplification was a 1.73-kb portion of the flagellin A gene of C. jejuni. The detection limit was lower than 30 cells per 100 ml in artificially contaminated waters. PCR assay and conventional culturing method had the same sensitivity, but results of the PCR technique were available within 48 h and so shortened the time necessary for detection by 48 h.
Hernandez
J
J
Departamento de Biotecnología, Universidad Politécnica, Valencia, Spain.
Alonso
J L
JL
Fayos
A
A
Amoros
I
I
Owen
R J
RJ
D001431:Bacteriological Techniques; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012623:Seawater; D012680:Sensitivity and Specificity; D014871:Water Microbiology
D017931:DNA Primers; D004269:DNA, Bacterial