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Abstract

Campylobacter bacteria are the causative agents of human foodborne acute enteric disease, ``campylobacteriosis''. Poultry meat is considered the primary source of this infection. This study was conducted to evaluate the use of multiplex polymerase chain reaction (multiplex PCR) for the direct detection of Campylobacter species from chilled chicken livers without an enrichment step, in comparison with the traditional culture-based method. Thirty chilled chicken liver samples were collected from supermarkets in Jazan City, representing six commercial poultry companies (A, B, C, D, E, and F). Samples were homogenized with buffered peptone water before being subjected to a multiplex PCR to detect the genus Campylobacter and three species: jejuni, lari, and coli, with PCR products of 816 bp, 323 bp, 251 bp, and 126 bp, respectively. In parallel, all samples were processed for the isolation of Campylobacter species using culture-based methods in accordance with ISO 10272-1:2017, using sterile Bolton Selective Enrichment Broth supplemented with 5% lysed blood and antibiotics, followed by plating on modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA). Only positive controls yielded PCR products, whereas no products were detected in the samples. Conversely, five samples (17%): S3 (from company A), S7 & S10 (from B), S18 (from D), and S23 (from E), showed bacterial growth on mCCDA plates. These five isolates were characterized by amplification and sequencing of the 16S rRNA gene. BLAST analysis in the NCBI database identified the four isolates (80%): S3, S7, S18, and S23 as E. coli, and the S10 isolate (20%) as Proteus mirabilis, indicating that culture-positive isolates were identified as non-Campylobacter species. The results suggested that direct detection of Campylobacter species using multiplex PCR could be a more reliable and rapid technique than the culture-based method, which primarily relies on an enrichment step and is time-consuming.

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