INTERPRETING CHROMOSOMAL DNA RESTRICTION PATTERNS PRODUCED BY PULSED-FIELD GEL-ELECTROPHORESIS - CRITERIA FOR BACTERIAL STRAIN TYPING
INTERPRETING CHROMOSOMAL DNA RESTRICTION PATTERNS PRODUCED BY PULSED-FIELD GEL-ELECTROPHORESIS - CRITERIA FOR BACTERIAL STRAIN TYPING
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DOI：
10.1128/jcm.33.9.2233-2239.1995
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发表时间：
1995-09-01
影响因子：
9.4
通讯作者：
SWAMINATHAN, B
中科院分区：
文献类型：
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作者：
TENOVER, FC;ARBEIT, RD;SWAMINATHAN, B
临床微生物学家经常被要求确定一组细菌分离株的亲缘关系，即对它们进行分型。在过去的十年中，传统的菌株分型方法，如噬菌体分型和血清分型，在许多实验室已经被较新的分子方法所补充或取代，如质粒指纹分析(43)、核糖体分型(40)、基于聚合酶链式反应的方法(45)和用脉冲场凝胶电泳法(PFGE)分析染色体DNA限制性内切酶图谱(4、14、27)。尽管噬菌体分型仍然被世界各地的一些大型参考实验室用于金黄色葡萄球菌的流行病学研究(36个)，血清分型仍然是沙门氏菌物种流行病学监测的有用工具(30个)，但需要一种可以用来对更广泛的细菌物种进行分型的菌株分型方法。目前，PFGE最接近于满足这一需求(3，42)。PFGE涉及将生物体嵌入琼脂糖凝胶中，原位裂解生物体，并用不频繁切割的限制性内切酶消化染色体DNA(14，27)。含有染色体DNA片段的琼脂糖切片被插入琼脂糖凝胶的孔中，并且通过根据预定图案切换电流方向的装置将限制片段分解为凝胶中的离散条带图案。然后将这些分离株的DNA限制性内切酶图谱相互比较，以确定它们的亲缘关系。目前，还没有用于分析片段模式的标准化标准。因此，对于哪些分离株应被指定为暴发相关菌株，哪些应被指定为非暴发相关菌株，不同的研究人员查看相同的PFGE结果可能会得出截然不同的结论。这篇嘉宾评论提出了一套解释PFGE产生的DNA限制模式的指导方针。作者是来自美国的研究人员，在过去几年里，他们将数十起疫情的流行病学数据与PFGE产生的菌株分型结果进行了关联。这些指南旨在供医院实验室的临床微生物学家使用，以检查相对
Clinical microbiologists are often asked to determine the relatedness of a group of bacterial isolates, that is, to type them. During the last decade, traditional methods of strain typing, such as bacteriophage typing and serotyping, have been supplemented or replaced in many laboratories with newer molecular methods, such as plasmid fingerprinting (43), ribotyping (40), PCR-based methods (45), and analysis of chromosomal DNA restriction patterns by pulsed-field gel electrophoresis (PFGE)(4, 14, 27). Although bacteriophage typing is still used in a number of large reference laboratories around the world for epidemiologic studies of Staphylococcus aureus (36) and serotyping continues to be a useful tool for epidemiologic surveillance of Salmonella species (30), there is a need for a method of strain typing that can be used to type a broader array of bacterial species. At present, PFGE comes closest to satisfying that need (3, 42).PFGE involves embedding organisms in agarose, lysing the organisms in situ, and digesting the chromosomal DNA with restriction endonucleases that cleave infrequently (14, 27). Slices of agarose containing the chromosomal DNA fragments are inserted into the wells of an agarose gel, and the restriction fragments are resolved into a pattern of discrete bands in the gel by an apparatus that switches the direction of current according to a predetermined pattern. The DNA restriction patterns of the isolates are then compared with one another to determine their relatedness. Currently, there are no standardized criteria for analyzing the fragment patterns. Consequently, different investigators viewing the same PFGE results may come to quite different conclusions as to which isolates should be designated as outbreak related and which should be designated as non-outbreak related. This guest commentary proposes a set of guidelines for interpreting DNA restriction patterns generated by PFGE. The authors are investigators from the United States who, over the last several years, have correlated epidemiologic data from dozens of outbreaks with strain typing results produced by PFGE. These guidelines are intended to be used by clinical microbiologists in hospital laboratories to examine relatively