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- Title
Rapid detection and enumeration of trh-carrying Vibrio parahaemolyticus with the alkaline phosphatase-labelled oligonucleotide probe.
- Authors
Raghunath, Pendru; Pradeep, Balakrishnan; Karunasagar, Indrani; Karunasagar, Iddya
- Abstract
An alkaline phosphatase (AP)-labelled oligonucleotide probe was developed to detect and enumerate trh+ Vibrio parahaemolyticus in seafood. The probe was evaluated using 40 isolates of V. parahaemolyticus, 45 isolates of other vibrios and 55 non-vibrio isolates. The probe reacted specifically with V. parahaemolyticus possessing either the trh1 or trh2 variant of the trh gene and was found to be 100% specific for trh+ V. parahaemolyticus. Using the trh probe, V. parahaemolyticus carrying trh gene was targeted in 34 seafood samples by direct plating and colony hybridization procedure. The trh+ V. parahaemolyticus could be detected in five of 34 (14.7%) samples and the levels ranged from 5.0 × 102 to 3.4 × 103 cfu g−1. Colonies of trh+ V.parahaemolyticus were isolated from the five positive samples. Forty seafood samples were analysed for trh+ V. parahaemolyticus by colony hybridization following enrichment in alkaline peptone water. 16 samples (40%) were positive for trh gene and trh+ V. parahaemolyticus was isolated from 15 samples (37.5%). To assess the sensitivity of the trh probe, seafood homogenates spiked with known concentrations of trh-positive V. parahaemolyticus were plated and hybridized. Counts obtained using the probe were similar to those of inocula. The results suggest that the AP-labelled trh probe is useful for the detection and enumeration of trh+ V. parahaemolyticus in seafood.
- Subjects
ALKALINE phosphatase; NUCLEIC acid probes; OLIGONUCLEOTIDES; MOLECULAR probes; SEAFOOD
- Publication
Environmental Microbiology, 2007, Vol 9, Issue 1, p266
- ISSN
1462-2912
- Publication type
Article
- DOI
10.1111/j.1462-2920.2006.01145.x