Detection of Common Molds and Yeasts in Low-Temperature Yogurt by TaqMan Probe Real-Time Fluorescence Quantitative Polymerase Chain Reaction
In order to improve the detection efficiency of molds and yeasts in low-temperature yogurt,primers and probes were designed according to the sequences of the conserved gene elongation factor-1α(EF-1α)for molds and yeasts,respectively.A real-time fluorescence quantitative polymerase chain reaction(real-time PCR)method with TaqMan probe was established for the detection of molds and yeasts,respectively.The genomic DNA of fungi and lactic acid bacteria commonly found in low-temperature yogurt,were used as amplification templates for specificity testing.The sensitivity and repeatability were also tested,and the standard curves were constructed.The results showed that the designed primers and probes had good specificity;the sensitivity of the method was 101 and 102 CFU/mL for yeasts and molds,respectively;the coefficients of variation within and between groups in the repeatability test were less than 2%,indicating good reliability and stability;the correlation coefficients of the standard curves were higher than 0.99.Therefore,the real-time PCR method enables the rapid detection of molds and yeasts in low-temperature yogurt.
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