title: Detection of Vibrio parahaemolyticus in shellfish using polymerase chain reaction-enzyme-linked immunosorbent assay.
abstract: OBJECTIVE
  This study evaluated the application of polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) for the detection of Vibrio parahaemolyticus in shellfish.
  
  
  RESULTS
  The PCRs were selected to amplify a species-specific sequence region. In particular, internal tl biotin-labelled oligonucleotide probe was used to capture the DIG-labelled PCR products. Next, the probe PCR product hybrids, immobilized on a streptavidin-coated microtiter plate, were detected with peroxidase-conjugated anti-digoxigenin antibody (anti-DIG-POD) and the colorimetric peroxidase substrate ABTS [2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)] using an ELISA plate reader.
  
  
  CONCLUSIONS
  The PCR-ELISA system described is a feasible, sensitive method for the direct and specific detection of V. parahaemolyticus in shellfish samples. Compared with gel-based detection methods, PCR-ELISA in this study increased sensitivity by 100-fold for V. parahaemolyticus.
  
  
  CONCLUSIONS
  The PCR-ELISA described may be used for potential rapid detection in routine shellfish analysis for the seafood industry. The sector requires simultaneous large-scale sample screenings to monitor contamination levels in processing plants and evaluate the performance of the hazard analysis and critical control point (HACCP) system. PCR-ELISA also proved to be economical, with a cost of about 9 Euros per sample, and the quick assay taking 8 h to complete starting from DNA extraction.
authors.affiliation: Dipartimento di Sanità Pubblica e Zootecnia, Facoltà di Medicina Veterinaria, Università degli Studi di Bari Aldo Moro, Valenzano, Bari, Italy. a.dipinto@veterinaria.uniba.it
authors.forename: A
authors.initials: A
authors.lastname: Di Pinto
authors.forename: V
authors.initials: V
authors.lastname: Terio
authors.forename: P
authors.initials: P
authors.lastname: Di Pinto
authors.forename: V
authors.initials: V
authors.lastname: Colao
authors.forename: G
authors.initials: G
authors.lastname: Tantillo
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis; D004076:Digoxigenin; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology / Q000379:methods*; D057230:Limit of Detection; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D017747:Seafood / Q000382:microbiology; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000382:microbiology*; D014736:Vibrio parahaemolyticus / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D004076:Digoxigenin