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Development of a PCR assay combined with a short enrichment culture for detection of Campylobacter jejuni in estuarine surface waters
Authors:Javier Hernandez  Jose L Alonso  Alicia Fayos  Inmaculada Amoros  robert J Owen
Affiliation:Departamento de Biotecnologia, Universidad Politécnica, Camino de Vera 14, 46022 Valencia, Spain; Institulo de Hidrología y Medio Natural, Universidad Politecnica, Camino de Vera 14, 46022 Valencia, Spain; National Collection of Type Cultures, Central Public Health Laboratory, 61 Colindale Ar., London NW9 5HT, UK
Abstract:
Abstract 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.
Keywords:Campylobacter jejuni    Polymerase chain reaction    DNA    Surface water
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