食品科学 ›› 2018, Vol. 39 ›› Issue (14): 166-171.doi: 10.7506/spkx1002-6630-201814025

• 生物工程 • 上一篇    下一篇

婴幼儿配方羊乳粉生产环节蜡样芽孢杆菌分离株多位点序列分型分析

刘阳1,葛武鹏1,*,张静1,郭春锋1,梁秀珍2,王智3,张兴吉1,王瑞1   

  1. (1.西北农林科技大学食品科学与工程学院,陕西?杨凌 712100;2.陕西省飞天乳业有限公司,陕西?宝鸡 721100;3.陕西百跃优利士乳业有限公司,陕西?咸阳 712000)
  • 出版日期:2018-07-25 发布日期:2018-07-16
  • 基金资助:
    陕西省战略性新兴产业重大产品(群)项目(2015KTCQ03-08;2016KTCQ03-03); 杨凌示范区科技攻关项目(2016NY-13)

Multiple Locus Sequence Typing Analysis of Bacillus cereus Isolates from the Production Line of Goat Milk-Based Infant Formula

LIU Yang1, GE Wupeng1,*, ZHANG Jing1, GUO Chunfeng1, LIANG Xiuzhen2, WANG Zhi3, ZHANG Xingji1, WANG Rui1   

  1. (1. College of Food Science and Engineering, Northwest A&F University, Yangling 712100, China; 2. Shaanxi Feitian Dairy Co. Ltd., Baoji 721100, China; 3. Shaanxi Baiyue Youlishi Dairy Co. Ltd., Xianyang 712000, China)
  • Online:2018-07-25 Published:2018-07-16

摘要: 为探讨我国婴幼儿配方羊乳粉生产环节阳性分离株的遗传多样性,并将其基因序列与已报道的乳源分离株的基因序列进行比对,确立其特征序列,为婴幼儿配方食品生产体系溯源提供依据,为探究其致病机理和有效防控提供参考依据。实验选取glpF、gmk、ilvD、pta、pur、pycA、tpi 7 个管家基因构建多位点序列分析分型方案,鉴定经随机扩增多态性DNA分型得到的42 株Bacillus cereus的序列类型。结果表明:42 株B. cereus分为7 个序列类型(sequence type,ST),分别为ST-770(4.8%,2/42)、ST-1000(71.4%,30/42)、ST-1084(9.5%,4/42)、ST-1348(2.4%,1/42)、ST-1349(2.4%,1/42)、ST-1350(2.4%,1/42)和ST-1351(7.1%,3/42);所有分离株被识别为205、142、23三个不同克隆谱系,2 个单态群(ST-770和ST-1351)和2 个独株(ST-1348和ST-1349);发现4 个新的ST型和4 个新的等位基因,已上传至国际数据库得到新的序列号和等位基因号,分别为ST-1348、ST-1349、ST-1350、ST-1351和glp-253、glp-254、ilv-277、pyc-199。

关键词: 蜡样芽孢杆菌(Bacillus cereus), 婴幼儿配方羊乳粉, 多位点序列分析分型(MLST), 基因序列(ST)

Abstract: In the present study, we analyzed the genetic diversity of Bacillus cereus isolated from the production line of goat milk-based infant formula, and we compared their gene sequences with published sequences of isolates from milk products, aiming to detect the specific sequence types of the isolates in this study, which will provide useful information for further research on pathogenesis, prevalence, traceability, prevention and control. A total of 350 B. cereus isolates were obtained in this study, which had been subtyped to 12 clusters (42 isolates) by random amplification of polymorphic DNA (RAPD). The multiple locus sequence typing (MLST) profiles for 42 isolates were conducted based on the house-keeping genes glpF, gmk, ilvD, pta, pur, pycA, and tpi. The clonal complexes for these isolates were analyzed by the online Burst (n-4). The results showed that 42 isolates were detected and classified into 7 sequence types: ST-770 (4.8%, 2/42), ST-1000 (71.4%, 30/42), ST-1084 (9.5%, 4/42), ST-1348 (2.4%, 1/42), ST-1349 (2.4%, 1/42), ST-1350 (2.4%, 1/42) and ST-1351 (7.1%, 3/42). Clonal complexes 205, 142, 23, and two groups (ST-770 and ST-1351) were detected as well as two singletons (3L33 with ST-1349 and 3L47 with ST-1350). Four novel STs (ST-1348, ST-1349, ST-1350, and ST-1351) and four novel alleles (glp-253, glp-254, ilv-277 and pyc-199) were found.

Key words: Bacillus cereus, goat milk-based infant formula, multiple locus sequence typing (MLST), sequence type (ST)

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