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DU Lixia, LI Dian, ZHUO Huirong, SHI Mengnan, CHEN Yanning, LI Lanfang, HOU Shaoyang. Primer probes for targeted identification and detection of Bifidobacterium animalis and their application[J]. Journal of China Pharmaceutical University, 2023, 54(2): 255-262. DOI: 10.11665/j.issn.1000-5048.20220912001
Citation: DU Lixia, LI Dian, ZHUO Huirong, SHI Mengnan, CHEN Yanning, LI Lanfang, HOU Shaoyang. Primer probes for targeted identification and detection of Bifidobacterium animalis and their application[J]. Journal of China Pharmaceutical University, 2023, 54(2): 255-262. DOI: 10.11665/j.issn.1000-5048.20220912001

Primer probes for targeted identification and detection of Bifidobacterium animalis and their application

Funds: This study was supported by the Natural Science Foundation of Shandong Province (No.ZR2022QB146); and the Heze University PhD Fund Planning (No.XY21BS35)
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  • Received Date: September 11, 2022
  • Revised Date: March 26, 2023
  • In this study, in order to overcome the shortcomings of the current methods used to identify Bifidobacterium animalis, such as long time, complicated operation and low adaptability of experimental environment, specific primer probes were designed based on ERIC-PCR technology to identify and detect B.animalis.Based on the genomic DNA of B.animalis HP-B1124, the ERIC-PCR reaction conditions of B.animalis HP-B1124 were optimized, and the ERIC-PCR fragments were obtained one by one and sequenced.Two pairs of specific primer probes were designed.The accuracy, specificity, limitation and universality of the two pairs of primer probes were evaluated, and the two pairs of specific primer probes were used for testing the products containing B.animalis in the commercially published formula.The two pairs of specific primer probes designed in this study could be used for identified strains of B.animalis more simply, quickly and targeted.This method has optimized the current relatively traditional methods of pure culture and plate counting identification of B.animalis, and has solved the high requirements of SNP genotyping technology and real-time fluorescence quantitative PCR method for experimental equipment and reagents in the identification of B.animalis to a certain extent.It has the characteristics of low cost, high specificity and earn a broad market development prospect.
  • [1]
    Sharples GJ, Lloyd RG. A novel repeated DNA sequence located in the intergenic regions of bacterial chromosomes[J]. Nucleic Acids Res, 1990, 18(22): 6503-6508.
    [2]
    Cao YF, Zhao LP. Computational analysis of the distribution of ERIC (IRU) in different bacterial genomes[J]. J Shanxi Univ Nat Sci Ed (山西大学学报 自然科学版), 2002, 25(4): 354-357.
    [3]
    Zhang ZC, Yang ES, Guo KK, et al. ERIC-PCR fingerprint analysis of intestinal flora of diarrhea piglets[J]. J Northwest A F Univ Nat Sci Ed (西北农林科技大学学报 自然科学版), 2019, 47(1): 31-38.
    [4]
    Hematzadeh A, Haghkhah M. Biotyping of isolates of Pseudomonas aeruginosa isolated from human infections by RAPD and ERIC-PCR[J]. Heliyon, 2021, 7(9): e07967.
    [5]
    Shrivastava UP. Molecular diversity assessment of PGPR using enterobacterial repetitive intergeneric consensus (ERIC) PCR[J]. Acad Voices, 2018, 7: 64-70.
    [6]
    Versalovic J, Koeuth T, Lupski JR. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes[J]. Nucleic Acids Res, 1991, 19(24): 6823-6831.
    [7]
    Tian M, Feng Z, Qu RD, et al. Identification of Chryseobacterium proteolyticum from different regions using ERIC-PCR and automated ribotyping[J]. Sci Technol Food Ind (食品工业科技), 2020, 41(1): 105-111, 118.
    [8]
    Chen YC, Cao YF, Zhao LP. Non-random nature of genomic DNA amplification of E.coli K-12 MG1655 via ERIC-PCR[J]. Microbiol China (微生物学通报), 2002, 29(6): 28-32.
    [9]
    Li P, Li JX, Li YF, et al. Analysis of ERIC-PCR fingerprints of Haemophilus parasuis field isolates from south-east of China[J]. Chin J Vet Sci (中国兽医学报), 2009, 29(12): 1566-1570.
    [10]
    Chen JC, Zhang JY, Zhan L, et al. Prevalence and antimicrobial-resistant characterization of Bacillus cereus isolated from ready-to-eat rice products in Eastern China[J]. Front Microbiol, 2022, 13: 964823.
    [11]
    Khoshbayan A, Golmoradi Zadeh R, Taati Moghadam M, et al. Molecular determination of O25b/ST131 clone type among extended spectrum β-lactamases production Escherichia coli recovering from urinary tract infection isolates[J]. Ann Clin Microbiol Antimicrob, 2022, 21(1): 35.
    [12]
    Zhang TF, Wang J, Yao Z, et al. Effect and mechanism of Bifidobacterium animalis B94 in the prevention and treatment of liver injury in rats[J]. Front Cell Infect Microbiol, 2022, 12: 914684.
    [13]
    Do MH, Oh MJ, Lee HB, et al. Bifidobacterium animalis ssp. lactis MG741 reduces body weight and ameliorates nonalcoholic fatty liver disease via improving the gut permeability and amelioration of inflammatory cytokines[J]. Nutrients, 2022, 14(9): 1965.
    [14]
    Lv XB, He LP, Zhang RJ, et al. Research progress in physiological functions of Bifidobacterium[J]. Sci Technol Food Ind (食品工业科技), 2013, 34(16): 353-358.
    [15]
    Liu H, Zhang KG, Liu P, et al. Improvement effect of Bifidobacterium animalis subsp. lactis MH-02 in patients receiving resection of colorectal polyps: a randomized, double-blind, placebo-controlled trial[J]. Front Immunol, 2022, 13: 940500.
    [16]
    Liu K, Kong XJ. Altered salivary microbiota following Bifidobacterium animalis subsp. Lactis BL-11 supplementation are associated with anthropometric growth and social behavior severity in individuals with prader-willi syndrome[J]. Probiotics Antimicrob Proteins, 2022, 14(4): 699-711.
    [17]
    Li L, Li JN, Yu WY. Brief introduction of the method of bacterial classification and identification[J]. J Anhui Agric Sci (安徽农业科学), 2004, 32(3): 549-551.
    [18]
    Deng MK, Sun Y, Han WQ. Methods of identification of bacteria[J]. Prog Biomed Eng (生物医学工程学进展), 2014, 35(2): 84-88.
    [19]
    Kim Y, Oh S, Park S, et al. Lactobacillus acidophilus reduces expression of enterohemorrhagic Escherichia coli O157: H7 virulence factors by inhibiting autoinducer-2-like activity[J]. Food Control, 2008, 19(11): 1042-1050.
    [20]
    Lomonaco S, Furumoto EJ, Loquasto JR, et al. Development of a rapid SNP-typing assay to differentiate Bifidobacterium animalis ssp. lactis strains used in probiotic-supplemented dairy products[J]. J Dairy Sci, 2015, 98(2): 804-812.
    [21]
    Zhao YP, Guo YW, Liu LF, et al. Determination of animal Bifidobacterium BB-12 in yogurt products by TaqMan-MGB fluorescent quantitative polymerase chain reaction[J]. J Food Saf Qual(食品安全质量检测学报), 2021, 12(24): 9362-9370.
    [22]
    Pang J, Liu ZY, Hao M, et al. Comparison and optimization of methods for genomic DNA extraction from Gram positive bacteria[J]. Microbiol China (微生物学通报), 2015, 42(12): 2482-2486.

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