食品中の原材料および食品添加物の迅速な検出法に関する研究

• Main Author: 鶴田 小百合
• Format: Thesis
• Language: Japanese
• Published: 三重大学 2015
• Subjects: Pacific; Theragra chalcogramma; Alaska
• Online Access: http://hdl.handle.net/10076/14585; https://mie-u.repo.nii.ac.jp/?action=repository_uri&item_id=10831; https://mie-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=10831&item_no=1&attribute_id=17&file_no=1

In this study, three assays for the rapid detection of ingredients and food additives, including fish roes, poisonous mushrooms, and sweeteners and preservatives in various foods, were developed to secure the safety and quality of food. In chapter 1, a rapid and sensitive TaqMan real-time polymerase chain reaction (PCR) assay was developed for the detection of Pacific cod (Gadus macrocephalus) and capelin (Mallotus villosus) roes in Alaska pollack (Theragra chalcogramma) roe products. Primers and TaqMan minor groove binder (MGB) probes were designed according to the gene encoding cytochrome b and evaluated by the specificity for 3 Alaska pollack roes, 3 Pacific cod roes, 9 capelin roes, and samples from 48 other species. Mitochondrial DNA extracts of all the target species had positive signals with no cross-reaction, the limit of detection being 0.002 ng/L of DNA. The linearity of the standard curves for the three species ranged from 0.002 ng/L to 20 ng/L of DNA with a square regression correlation (R2) of 1.000. This assay was applied for the detection of Pacific cod and capelin roes in mixture samples: Pacific cod or capelin roes were added to Alaska pollack roes at 0.1%, 1%, and 10%. The threshold cycle (Ct) values were obtained from both the mixture samples at 0.1%. The practical applicability of this assay was examined in 64 samples of Alaska pollack roe products. In all cases, the species detected from the samples corresponded with the species described on the food label. This new assay proved to be specific to the target species, highly sensitive, and applicable tofood samples, making it useful for the rapid detection of Pacific cod and capelin roes in Alaska pollack roe products. In chapter 2, a rapid multiplex real-time PCR assay was developed to achieve highly specific, simultaneous detection of two types of mushrooms: the poisonous mushroom Omphalotus guepiniformis and the edible mushroom Lentinula edodes, both of which have fruiting bodies with a similar morphology. Primers and TaqMan MGB probes ...