Journal of Dali University ›› 2019, Vol. 4 ›› Issue (2): 33-.

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Establishment of a Real-time PCR Method for Detection of Gut Bacteria in Patients with Ulcerative Colitis

Duan Xiaomei, Li Chaonan, Geng Funeng, Liu Bin, Zhao Yu, Li Yue   

  1. （1. Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D, Dali, Yunnan 671000, China；
    2. National-Local Joint Engineering Research Center of Entomoceutics, Dali, Yunnan 671000, China；3.Yunnan Provincial
    Collaborative Innovation Center for Entomoceutics, Dali , Yunnan 671000, China; 4. Key Laboratory of
    Periplaneta americana in Sichuan Province, Chengdu 610000, China）
  • Received:2018-10-31 Revised:2018-12-23 Online:2019-02-15 Published:2019-02-15

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Abstract: 〔Abstract〕Objective: To establish a quantitative real-time PCR method for detecting gut bacteria in patients with ulcerative colitis
in order to further explore the effects of imbalanced gut flora on ulcerative colitis（UC）. Methods: We identified 7 gut microbes that
were strongly associated with the pathogenesis of UC. Specific PCR primers were designed to construct a standard recombinant plasmid
containing specific amplified fragments. The absolute quantitative method of real-time PCR was established by using recombinant
plasmids as standards. Results: The real-time PCR method for the detection of such target bacteria as Bifidobacterium, Escherichia
coli, and total intestinal flora, was successfully established. The standard curve of gut bacteria showed a good linear relation with
correlation coefficient ≥0.998. Conclusion: The SYBR Green real-time PCR method can detect seven intestinal bacteria in ulcerative
colitis specifically and provide a convenient and effective method for exploring the correlation between the imbalance of gut bacteria
and UC pathogenesis.

Key words: 〔Key words〕Real-time PCR, gut bacteria, Ulcerative Colitis recombinant plasmid