Establishment and Application of TaqMan Probe Real-time Fluorescent PCR Detection Method for Blueberry Honey in Guizhou
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Abstract:
In this study, a highly efficient and accurate method was developed for identifying Guizhou characteristic blueberry honey (from honey bee Apis cerana) based on specific TaqMan probe of real-time fluorescent PCR. Blueberry honey samples (including Apis mellifera) were collected from the blueberry growing areas of Bai zhulin, Maka and Wukaping in Guizhou, meanwhilw, commercial local honey and Canadian blueberry honey were also purchased. Twenty-six blueberry samples at the same flowering stage were collected from the blueberry gardens and the areas around the bee farms in the Baizhulin region of Majiang Town, Guizhou Province. The trnL gene in the sampling plant genome were sequenced. A trnL gene-specific primer of blueberry was designed based on the multiple sequence alignments of trnL gene sequences in the plant genome. TaqMan probe was used for validation. The results showed that the TaqMan probe had high specificity, and the minimum concentration of DNA as low as 0.3 ng/μL can be detected was in blueberry sample using the established TaqMan probe real-time fluorescent PCR detection method. Such an established method was used to test the samples of 11 kinds of commercial honey and Guizhou blueberry honey. It was found that the Ct value of Guizhou blueberry honey was 24~26, the number of blueberry pollen was 800~1 700, and the Ct values of other honey samples were over 30, indicating that this method can effectively distinguish Guizhou blueberry honey from other honeys, thereby having a good application prospect.
