CAJ | 학술논문

目的建立一种可共享的猪链球菌脉冲场凝胶电泳分析(PFGE)方法.方法利用PulseNet技术优化猪链球菌PFGE分子分型方法,提出新的分析方案,包括胶块的制备、内切酶的选择、电泳条件等.利用DNAStar的Mapdraw软件对猪链球菌基因组序列进行分析,发现在242种内切酶中,Swa Ⅰ、Sma Ⅰ、Apa Ⅰ种酶比较适合.结果通过对34个血清型共100株菌的分析,发现使用Swa Ⅰ可获得59种带型,使用Sma Ⅰ可获得53种带型,使用Apa Ⅰ可获得43种带型.其中以Swa Ⅰ的分辨率最好.结论提出猪链球菌的Swa Ⅰ PFGE分析方法,并对条件进行优化,较原方法所需时间短、图像清晰,更具有可重复性,基本具备推广的条件.
목적 건립일충가공향적저련구균맥충장응효전영분석(PFGE)방법.방법 이용PulseNet기술우화저련구균PFGE분자분형방법,제출신적분석방안,포괄효괴적제비、내절매적선택、전영조건등.이용DNAStar적Mapdraw연건대저련구균기인조서렬진행분석,발현재242충내절매중,Swa Ⅰ、Sma Ⅰ、Apa Ⅰ충매비교괄합.결과 통과대34개혈청형공100주균적분석,발현사용Swa Ⅰ가획득59충대형,사용Sma Ⅰ가획득53충대형,사용Apa Ⅰ가획득43충대형.기중이Swa Ⅰ적분변솔최호.결론 제출저련구균적Swa Ⅰ PFGE분석방법,병대조건진행우화,교원방법소수시간단、도상청석,경구유가중복성,기본구비추엄적조건.
Objective To develop a PFGE protocol for Streptococcus suis.Methods We developed and optimized a PFGE protocol for S.suis,in terms of plug preparation,choice of restriction endonucleases and optimized electrophoresis parameters.By analyzing the genome sequences of S.suis P1/7 with Mapdraw of DNAStar.we found three restriction enzymes,Swa Ⅰ,Sma Ⅰ and Apa Ⅰ,were more suitable than others.Results Analysis of 100 isolates of S.suis including 34 of 35 serotypes identified,59,53 and 43 patterns were obtained from Swa Ⅰ,Sma Ⅰ and Apa Ⅰ restriction,respectively.The enzyme Swa Ⅰ had the greatest power for discrimination ability.Conclusion By optimization of the protocol at various conditions,a rapid,reproducible,economic and practical PFGE method for S.suis was developed.