The establishment of a novel method of nano-immunomagnetic separation and Real-time PCR for detecting Vibrio cholerae from seafood / 中华预防医学杂志
Chinese Journal of Preventive Medicine
; (12): 133-137, 2014.
Article
em Zh
| WPRIM
| ID: wpr-298954
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WPRO
ABSTRACT
<p><b>OBJECTIVE</b>A novel method of Nano-Immunomagnetic Separation (Nano-IMS) plus Real-time PCR was established for detecting Vibrio cholerae.</p><p><b>METHODS</b>The Nano-Immunomagnetic Beads were created by using the monoclonal antibody of Vibrio cholerae, which was named Nano-IMB-Vc. Nano-IMB-Vc has specific adsorption of Vibrio cholerae, combined with Real-time PCR technology, a method for rapid detection of Vibrio cholerae was established. The capture specificity of Nano-IMB-Vc was tested by using 15 bacteria strains. The specificity of Real-time PCR method was tested by using 102 targets and 101 non-targets bacteria strains. The sensitivity of Nano-IMS plus Real-time PCR were tested in pure culture and in artificial samples and compared with NMKL No.156.</p><p><b>RESULTS</b>The capture ratio of Nano-IMB-Vc was reached 70.2% at the level of 10(3) CFU/ml. In pure culture, the sensitivity of Nano-IMS plus Real-time PCR was reached at 5.4×10(2) CFU/ml. The specific of Real-time PCR method was tested by using 102 targets and 101 non-targets bacteria. The results showed that 102 strains of Vibrio cholerae test results were all positive, and the rest of the 101 strains of non-target bacteria test results were negative. No cross-reaction was founded. Add 1 CFU vibrio cholerae per 25 g sample, it could be detect with Nano-IMS plus Real-time PCR method after 8 hours enrichment.</p><p><b>CONCLUSIONS</b>The Nano-IMS plus Real-time PCR method of Vibrio cholerae established in this study has good specificity and sensitivity, which could be applied to the rapid detection of Vibrio cholerae.</p>
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Assunto principal:
Vibrio cholerae
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Vibrio parahaemolyticus
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Alimentos Marinhos
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Separação Imunomagnética
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Nanotecnologia
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Reação em Cadeia da Polimerase em Tempo Real
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Microbiologia de Alimentos
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Genética
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Métodos
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Microbiologia
Idioma:
Zh
Revista:
Chinese Journal of Preventive Medicine
Ano de publicação:
2014
Tipo de documento:
Article