Study on LAMP for Detecting Gluconobacter in Yogurt
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Abstract:
Objective: The aim was to develop a rapid, simple, and sensitive LAMP method for the detection of Gluconobacter to be used in the yogurt industry. Method: Gluconobacter oxydans was selected as the type culture strain. Two pairs of LAMP primers targeting the 16S rRNA（X73820）housekeeping gene of Gluconobacter available in GenBank were designed. Concentrations of the primers and Mg2+ and the temperature of the LAMP were optimized, and a rapid, sensitive LAMP assay for Gluconobacter detection was established. Sensitivity of the assay was evaluated and compared with the PCR assay. Results: It was found that the LAMP primers showed high specificity and the LAMP product sizes, after restriction enzyme digestion, were consistent with the theoretical fragment sizes. The sensitivity of the LAMP assay for Gluconobacter was 7.5 × 101 CFU/mL. The LAMP assay was found to be 10-fold more sensitive than the traditional PCR assay. The LAMP detection limit was 7.5 × 102 CFU/mL for artificial contaminated yogurt. Conclusion: LAMP amplification established in this study provides a rapid,highly specific and sensitive method for Gluconobacter detection. The LAMP method is valuable for its application and exploitation in food industries.
