COMPARISON OF RIBOTYPING AND MULTILOCUS ENZYME ELECTROPHORESIS FOR SUBTYPING OF LISTERIA-MONOCYTOGENES ISOLATES
COMPARISON OF RIBOTYPING AND MULTILOCUS ENZYME ELECTROPHORESIS FOR SUBTYPING OF LISTERIA-MONOCYTOGENES ISOLATES
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DOI：
10.1128/jcm.32.12.2936-2943.1994
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发表时间：
1994-12-01
影响因子：
9.4
通讯作者：
SCHUCHAT, A
中科院分区：
文献类型：
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作者：
GRAVES, LM;SWAMINATHAN, B;SCHUCHAT, A
对临床和非临床来源的305株单核细胞增多性李斯特菌进行核糖体分型和多位点酶电泳法的亚型比较。用琼脂糖凝胶电泳法分离单核细胞增多性李斯特菌的EcoRI限制性片段，并用地高辛标记的克隆的大肠杆菌rrnB操纵子(PKK3535)进行Southern杂交。单核细胞增多性李斯特菌分离株分为28个不同的核糖核型，而MEE分析将同一分离株分为78个电泳型(ETS)。根据其核型图谱，这些菌株被分为两个亚组。核糖型α(RTα)亚群含有血清型1/2a。1/2c和3a，核糖型β(RTP)亚群包括1/2b、3b、4b和4ab型。这种划分与MEE分析完全一致，MEE分析将该物种分为两个亚群(ET群A和B)，每个亚群的血清型分布相同。总体而言，MEE比核糖体测序更具鉴别力。然而，在一些情况下，核糖体分型可以区分同一ET内的不同分离株。核糖体分型对血清型1/2a、1/2c和3a(Simpson‘s多样性指数[DI]=0.81)的鉴别率高于对1/2b和4b型(DI=0.76)。相当大比例(69%)的血清1/2b和4b型菌株聚集在5个ETS和5个核型中。这些数据表明，核糖体分型和MEE不能充分区分血清1/2b和4b型菌株。应探索脉冲场凝胶电泳法和随机扩增多态性DNA分析等方法进一步鉴别这些血清型菌株。
Ribotyping was compared with multilocus enzyme electrophoresis (MEE) for subtyping 305 Listeria monocytogenes isolates from clinical and nonclinical sources. For ribotyping, EcoRI-restricted genomic DNA fragments of L. monocytogenes strains were separated by agarose gel electrophoresis, and Southern blots were probed with a cloned Escherichia coli rrnB operon (plasmid pKK3535) labeled with digoxigenin. The L. monocytogenes isolates were divided into 28 distinct ribotypes, while MEE analysis divided the same isolates into 78 electrophoretic types (ETs). On the basis of their ribotype profiles, the strains were divided into two subgroups. The ribotype alpha (RT alpha) subgroup contained serotypes 1/2a. 1/2c, and 3a, and the ribotype beta (RTP) subgroup contained serotypes 1/2b, 3b, 4b, and 4ab. This division is in complete agreement with MEE analysis, which divides the species into two subgroups (ET groups A and B), with the same serotype distribution in each subgroup. Overall, MEE was more discriminating than ribotyping. However, in several instances ribotyping discriminated between isolates within the same ET. Ribotyping was more discriminating for serotypes 1/2a, 1/2c, and 3a (Simpson's Index for Diversity [DI] = 0.81) than for serotypes 1/2b and 4b (DI = 0.76). A substantial proportion (69%) of serotype 1/2b and 4b strains clustered in five ETs and five ribotypes. These data suggest that ribotyping and MEE do not provide adequate discrimination between strains of serotypes 1/2b and 4b. Methods such as pulsed-field gel electrophoresis and random amplified polymorphic DNA analysis should be explored for further discrimination of strains of these serotypes.