Detection of Verocytotoxygenic Escherichia coli (VTEC) from Brilliant Green Lactose Bilebroth (BGLB) by Polymerase Chain Reaction (PCR)

摘要

Verocytotoxygenic Escherichia coli (VTEC) is pathogenic bacteria for human and associated with dairy milk and dairy farm environment. VTEC isolation were carried out by inoculating in BGLB as enrichment media and then specified in Eosin Methylene Blue (EMB) agar. This study aim to identify VTEC isolated from BGLB using PCR method. Six VTEC sample were used in this study, and those samples were collected from: milk bulktank (n=1), hand swab before milking (n=1), and hand swab after milking (n=1), swab of milk filter (n=1), feces (n=1), and water from dairy drinking places (n=1). All samples were inoculated in BGLB, and then bacterial DNA were extracted. PCR amplification using specific primer of vt_1 and vt_2 targeting on.....gene and....gene, with expected band at 130 bp and 779 bp, respectively. Positive results were sequenced and then analyzed using BLAST. Our results showed two out of six were positive of VTEC.