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Articles

Development of an indirect enzyme-linked immunosorbent assay for the detection of osmotolerant yeast Zygosaccharomyces rouxii in different food

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Pages 976-988 | Received 24 Mar 2018, Accepted 23 May 2018, Published online: 08 Oct 2018
 

ABSTRACT

A polyclonal antibody-based indirect ELISA for the sensitive and specific detection of osmotolerant Zygosaccharomyces rouxii was established. The reactivity of the polyclonal antibodies (pAbs) with Z. rouxii was confirmed by immunofluorescence. The ELISA detection limit was 5 × 104 CFU/mL. For an evaluation of the specificity of pAbs, reactivity to 10 fungi species and 2 bacteria species commonly found in food environments was analysed by indirect ELISA. ELISA reacts with all tested Z. rouxii, whereas no cross-reactions or weakly reactions with 9 fungi and 2 bacteria were observed. The immunoassay staining confirmed the ELISA. Further on, the assay applicability was tested with concentrated apple juice, wine, and honey. Food samples were inoculated with 100 CFU/mL of Z. rouxii. After enrichment for 24 h, the established ELISA can successfully detect contaminated wine and concentrated apple juice samples, while only 30 h was required for honey samples.

Acknowledgements

The authors would like to acknowledge Mrs Wang, who is responsible for the Life science research platform laser confocal chamber of Northwest A & F University. She generously provided the microscopy technology.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by National Natural Science Foundation of China [31371814]; major projects of strategic emerging industries in Shaanxi province of China [2016KTCQ03-12].