Comparison of different primers for rapid detection of Vibrio parahaemolyticus using the polymerase chain reaction

Luan, Xiaodong; Chen, J.-X.; Zhang, X.-H.; Jia, Jingdun; Sun, Feng-Xian; Li, Y.

doi:10.1111/j.1472-765x.2006.02074.x

Cited by 31 publications

(26 citation statements)

References 13 publications

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“…Polymerase chain reactions were used to confirm the identity of colonies on BHI-NaCl plates. Colonies of V. parahaemolyticus were confirmed by using VPM1 and VPM2 primers specific for the metalloprotease gene (Luan et al, 2007). Vibrio vulnificus was confirmed by using Cyt1 and Cyt2 primers targeting the haemolysin/cytolysin gene (Morris et al, 1987).…”

Section: Pcr Verification Of Vibriosmentioning

confidence: 99%

Light‐scattering sensor for real‐time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate

Huff

Aroonnual

Littlejohn

et al. 2012

Microbial Biotechnology

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SummaryThe three most common pathogenic species of Vibrio, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus, are of major concerns due to increased incidence of water‐ and seafood‐related outbreaks and illness worldwide. Current methods are lengthy and require biochemical and molecular confirmation. A novel label‐free forward light‐scattering sensor was developed to detect and identify colonies of these three pathogens in real time in the presence of other vibrios in food or water samples. Vibrio colonies grown on agar plates were illuminated by a 635 nm laser beam and scatter‐image signatures were acquired using a CCD (charge‐coupled device) camera in an automated BARDOT (BActerial Rapid Detection using Optical light‐scattering Technology) system. Although a limited number of Vibrio species was tested, each produced a unique light‐scattering signature that is consistent from colony to colony. Subsequently a pattern recognition system analysing the collected light‐scatter information provided classification in 1−2 min with an accuracy of 99%. The light‐scattering signatures were unaffected by subjecting the bacteria to physiological stressors: osmotic imbalance, acid, heat and recovery from a viable but non‐culturable state. Furthermore, employing a standard sample enrichment in alkaline peptone water for 6 h followed by plating on selective thiosulphate citrate bile salts sucrose agar at 30°C for ∼ 12 h, the light‐scattering sensor successfully detected V. cholerae, V. parahaemolyticus and V. vulnificus present in oyster or water samples in 18 h even in the presence of other vibrios or other bacteria, indicating the suitability of the sensor as a powerful screening tool for pathogens on agar plates.

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Section: Pcr Verification Of Vibriosmentioning

confidence: 99%

Light‐scattering sensor for real‐time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate

Huff

Aroonnual

Littlejohn

et al. 2012

Microbial Biotechnology

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“…(1996) to differentiate A. salmonicida subspecies salmonicida from other A. salmonicida subspecies. Other bacterial species used in this work were also subjected to PCR confirmation (Khan & Cerniglia 1997; Urdaci, Chakroun, Faure & Bernardet 1998; LeJeune & Rurangirwa 2000; Scarpellini, Franzetti & Galli 2004; Xia, Ma, Rahman & Wu 2004; Luan, Chen, Zhang, Jia, Sun & Li 2007; Nam & Joh 2007).…”

Section: Methodsmentioning

confidence: 99%

Antibody-coated gold nanoparticles immunoassay for direct detection of Aeromonas salmonicida in fish tissues

Saleh

Soliman

Haenen

et al. 2011

Journal of Fish Diseases

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Aeromonas salmonicida is the causative agent of furunculosis, a disease that affects both salmonid and non-salmonid fish. Detection of A. salmonicida can be labour intensive and time consuming because of the difficulties in distinguishing the bacterium from other species given the wide variety of existing biochemical profiles and the slow growth characteristics which allow other organisms to overgrow the A. salmonicida. Herein, we report the development of a specific immunoassay using gold-conjugated polyclonal antibodies for the rapid detection of A. salmonicida in fish tissues. Monodispersible 13-nm gold nanoparticles were coated with polyclonal antibodies specific to A. salmonicida. Reddish purple agglutination of gold particles indicated the presence of A. salmonicida in samples. Positive reactions were detected visually with the naked eye. No agglutination was observed when A. salmonicida antibody-coated gold nanoparticles were tested with other common bacterial fish pathogens, thereby verifying the specificity of the assay. The assay could detect A. salmonicida in fish tissues down to 1 × 10(4) CFU mL(-1) , and results were obtained within 45 min. The antibody-coated gold nanoparticles were stable for at least 2 months at 4 ° C. The immunoassay using antibody-coated gold nanoparticles represents a promising tool for the rapid and specific detection of A. salmonicida in fish tissues.

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“…While primers amplifying toxR have been used by most investigators for detection of total V. parahaemolyticus, few have used primers amplifying gyrB, tlh, and vpm gene encoding a metalloprotease or V. parahaemolyticus sequence in a recombinant plasmid pR72 H [17,18]. PCR amplification of toxR gene in lysates of enrichment broths at 6 hours detected V. parahaemolyticus in a larger number of samples compared to the conventional culture method [19].…”

Section: Detection Of Bacterial Pathogensmentioning

confidence: 99%

Molecular Detection of Pathogens in Seafood

Karunasagar

2010

Handbook of Seafood Quality, Safety and Health Applications

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Comparison of different primers for rapid detection of Vibrio parahaemolyticus using the polymerase chain reaction

Cited by 31 publications

References 13 publications

Light‐scattering sensor for real‐time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate

Light‐scattering sensor for real‐time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate

Antibody-coated gold nanoparticles immunoassay for direct detection of Aeromonas salmonicida in fish tissues

Molecular Detection of Pathogens in Seafood

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