中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2011年
12期
1086-1089
,共4页
董银苹%崔生辉%于红霞%李凤琴
董銀蘋%崔生輝%于紅霞%李鳳琴
동은평%최생휘%우홍하%리봉금
乳酸杆菌属%链球菌,嗜热%细菌分型技术%电泳,凝胶,脉冲场
乳痠桿菌屬%鏈毬菌,嗜熱%細菌分型技術%電泳,凝膠,脈遲場
유산간균속%련구균,기열%세균분형기술%전영,응효,맥충장
Lactobacillus%Streptococcus thermophilus%Bacterial typing techniques%Electrophoresis,gel,pulsed-field
目的 建立乳酸杆菌及嗜热链球菌的PFGE分子分型方法,并对北京市售酸奶中分离的乳酸杆菌及嗜热链球菌进行分子分型.方法 选取ApaⅠ、NotⅠ、sfiⅠ、XbaⅠ和SmaⅠ共5种PFGE分析中常用的限制性内切酶,对从北京市售酸奶中分离到的52株乳酸杆菌、嗜热链球菌以及相应的标准菌株进行酶切,优化PFGE限制性内切酶种类及电泳条件,并用优化出的实验条件对菌株进行分子分型,同时进行聚类分析,与生化鉴定及16s rRNA基因鉴定结果进行对比分析.结果 限制性内切酶NotⅠ对保加利亚乳酸杆菌、发酵乳酸杆菌和德氏乳酸杆菌的酶切效果较好,而限制性内切酶Apa Ⅰ对嗜热链球菌、嗜酸乳酸杆菌及干酪乳酸杆菌的酶切效果较好.24株保加利亚乳酸杆菌被分为8个PFGE型,15株嗜热链球菌被分为8个PFGE型,7株嗜酸乳酸杆菌被分为3个PFGE型,2株德氏乳酸杆菌分属于2个不同的PFGE型.结论 建立的PFGE方法分析结果与生化鉴定及16s rRNA基因鉴定结果高度符合,所建方法适用于乳酸杆菌及嗜热链球菌的分子分型.
目的 建立乳痠桿菌及嗜熱鏈毬菌的PFGE分子分型方法,併對北京市售痠奶中分離的乳痠桿菌及嗜熱鏈毬菌進行分子分型.方法 選取ApaⅠ、NotⅠ、sfiⅠ、XbaⅠ和SmaⅠ共5種PFGE分析中常用的限製性內切酶,對從北京市售痠奶中分離到的52株乳痠桿菌、嗜熱鏈毬菌以及相應的標準菌株進行酶切,優化PFGE限製性內切酶種類及電泳條件,併用優化齣的實驗條件對菌株進行分子分型,同時進行聚類分析,與生化鑒定及16s rRNA基因鑒定結果進行對比分析.結果 限製性內切酶NotⅠ對保加利亞乳痠桿菌、髮酵乳痠桿菌和德氏乳痠桿菌的酶切效果較好,而限製性內切酶Apa Ⅰ對嗜熱鏈毬菌、嗜痠乳痠桿菌及榦酪乳痠桿菌的酶切效果較好.24株保加利亞乳痠桿菌被分為8箇PFGE型,15株嗜熱鏈毬菌被分為8箇PFGE型,7株嗜痠乳痠桿菌被分為3箇PFGE型,2株德氏乳痠桿菌分屬于2箇不同的PFGE型.結論 建立的PFGE方法分析結果與生化鑒定及16s rRNA基因鑒定結果高度符閤,所建方法適用于乳痠桿菌及嗜熱鏈毬菌的分子分型.
목적 건립유산간균급기열련구균적PFGE분자분형방법,병대북경시수산내중분리적유산간균급기열련구균진행분자분형.방법 선취ApaⅠ、NotⅠ、sfiⅠ、XbaⅠ화SmaⅠ공5충PFGE분석중상용적한제성내절매,대종북경시수산내중분리도적52주유산간균、기열련구균이급상응적표준균주진행매절,우화PFGE한제성내절매충류급전영조건,병용우화출적실험조건대균주진행분자분형,동시진행취류분석,여생화감정급16s rRNA기인감정결과진행대비분석.결과 한제성내절매NotⅠ대보가리아유산간균、발효유산간균화덕씨유산간균적매절효과교호,이한제성내절매Apa Ⅰ대기열련구균、기산유산간균급간락유산간균적매절효과교호.24주보가리아유산간균피분위8개PFGE형,15주기열련구균피분위8개PFGE형,7주기산유산간균피분위3개PFGE형,2주덕씨유산간균분속우2개불동적PFGE형.결론 건립적PFGE방법분석결과여생화감정급16s rRNA기인감정결과고도부합,소건방법괄용우유산간균급기열련구균적분자분형.
Objective To develop a pulsed field gel electrophoresis (PFGE) method for molecular typing of Lactobacillus and Streptococcus thermophilus (S.thermophilus) and to apply it in identification and characterization of both bacteria isolated from yoghourt collected from Beijing supermarket.Methods The five most useful restriction enzymes includingApa Ⅰ,Not ],Sfi Ⅰ,Xba Ⅰ and Sma Ⅰ were chosen to cut DNA of 52 strains of Lactobacillus,S.thermophilus as well as associated standard bacteria strains.The endonucleases and electrophoresis conditions for PFGE analysis were optimized and applied in molecular typing of Lactobacillus and S.thermophilus isolates.Cluster analysis based on the PFGE data was conducted.The identification results of PFGE were compared with those obtained in biochemical and 16s ribosomal RNA PCR identification tests.Results Not Ⅰ was suitable for L.bulgaricus,L.fermentum and L.delbrueckii digestion.While Apa Ⅰ was an appropriate endonuclease for S.thermophilus,L.acidophilus and L.casei digestion.The results of molecular typing indicated that 24 strains of L.bulgaricus and 15 strains of S.thermophilus were grouped into 8 types by PFGE method,respectively.While 7 strains of L.acidophilus were grouped into 3 types and 2 strains of L.delbrueckii were grouped into 2 different PFGE types.Conclusion The results of PFGE analysis are in compliance with those of 16s rRNA PCR and biochemical identification.The PFGE method developed in this study is suitable for molecular characterization of both Lactobacillus and S.thermophilus.
