AP-PCR基因指纹用于变形链球菌传播株与非传播株的筛选
Detection of the Transmitted Strains and Non-transmitted Strains ofMutans Streptococciby AP-PCR
作者：李颂,刘天佳,庄姮,杨锦波,杨德琴,刘昭慧
Author：LISong,LIUTianjia,ZHUANGHeng,et al
收稿日期：2003-10-25 年卷（期）页码：2003,21(05):392-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：变形链球菌,传播,随机引物—聚合酶链反应,基因型,定植,
Key words：mutans streptococci,transmission,genotype,colonization,AP-PCR(arbitrarily primed polymerase chain reaction),
基金项目：
本课题为国家自然科学基金(编号30171013)和安徽省教育厅基金(2000JI125)资助项目
中文摘要
目的 应用AP-PCR基因指纹筛选变形链球菌(mutans streptococci,MS)的传播株(transmitted strains)与非传播株(nontransmitted strains),探讨影响MS传播的因素。方法 选取20对口腔中已定居有MS的儿童(3~4岁) 与他们的母亲,取牙面菌斑样本涂于轻唾—杆菌肽培养基。每人随机挑取45株分离株,提取染色体DNA,AP-PCR 基因指纹检测。结果 ①从200个分离株中共分辨出45个不同的基因型,其中10位(50%)母亲和15位(75%)儿童分别带有1种类型,另5位(25%)儿童带2种类型,另10位(50%)母亲带有2种或2种以上类型(2位母亲带有5 种型),表明人群口腔中定居的MS存在基因多态性;②比较母亲与其子女MS基因型的相似性发现,20对母子中 16对(80%)有相似基因型出现,提示MS在此人群中的高传播现象;③对有传播现象的16位母亲的S.mutans进行传播株与非传播株的筛选发现,10(50%)位母亲口腔中传播株与非传播株共存,表明并非所有基因型的S.mutans 都能传播。结论 ①AP-PCR基因指纹能清晰地分辨出S.mutans的传播株和非传播株;②在S.mutans的母婴传播过程中某一型菌株的优先传播是普遍存在的,进一步探讨造成这种现象的原因是很有必要的。
英文摘要
Objective Dental caries is a transmissible infectious disease in whichS.mutansplays the major role. The purpose of this study was to detect theS.mutanstransmitted strains and non-transmitted strains by AP-PCR fingerprint for laying the foundation of study on the relation between bacterial properties ofS.mutansand its transmission.Methods Plaque samples were obtained from buccal surfaces of 20 3~4 years old children and their mothers. Caries experience was scored by dmft (DM- FT). Diet regime, breast feeding and care of the childrenwere recorded. 200 Isolates ofS.mutanswere detected byS.mutansB medium and confirmed biochemically. DNA from each isolate was purified and AP-PCR fingerprinting was conducted. Amplicons were separated by electrophoresis in 1·5% agarose gels.Results 45 different patterns among the 200 isolateswere found. There were 10 mothers (50%) and 15(75%) children owning one genotype while 10 mothers and 5 children owning more than one (2 mothers owning 5 types). The data showed that the mothers harbored a more heterogeneous population ofS.mutansthan their children. Comparisons in genotypes between children and theirmothers discovered that 16(80%)children harbored the same geno- types as their mothers, indicating hightransmission in the group of people. Detection of theS.mutanstransmitted strains and non- transmitted strains in mothers demonstrated that 10 (50%) mothers harbored not only transmitted strains but also non-transmitted strains, suggesting that different strains had different ability of transmission.Conclusion AP-PCR was capable of detecting the S.mutanstransmitted strains and non-transmitted strains. SomeS.mutansgenotypes had higher ability of transmission than oth- ers.
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