Sander A, Ruess M, Bereswill S, Schuppler M, Steinbrueckner B
Abteilung Mikrobiologie und Hygiene, Institut für Medizinische Mikrobiologie und Hygiene, Klinikum der Universität Freiburg, Freiburg, Germany.
J Clin Microbiol. 1998 Oct;36(10):2973-81. doi: 10.1128/JCM.36.10.2973-2981.1998.
Seventeen isolates of Bartonella henselae from the region of Freiburg, Germany, obtained from blood cultures of domestic cats, were examined for their genetic heterogeneity. On the basis of different DNA fingerprinting methods, including pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus (ERIC)-PCR, repetitive extragenic palindromic (REP) PCR, and arbitrarily primed (AP)-PCR, three different variants were identified among the isolates (variants I to III). Variant I included 6 strains, variant II included 10 strains, and variant III included only one strain. By all methods used, the isolates could be clearly distinguished from the type strain, Houston-1, which was designated variant IV. A previously published type-specific amplification of 16S rDNA differentiated two types of the B. henselae isolates (16S rRNA types 1 and 2). The majority of the isolates (16 of 17), including all variants I and II, were 16S rRNA type 2. Only one isolate (variant III) and the Houston-1 strain (variant IV) comprised the 16S rRNA type 1. Comparison of the 16S rDNA sequences from one representative strain from each of the three variants (I to III) confirmed the results obtained by 16S rRNA type-specific PCR. The sequences from variant I and variant II were identical, whereas the sequence of variant III differed in three positions. All methods applied in this study allowed subtyping of the isolates. PFGE and ERIC-PCR provided the highest discriminatory potential for subtyping B. henselae strains, whereas AP-PCR with the M13 primer showed a very clear differentiation between the four variants. Our results suggest that the genetic heterogeneity of B. henselae strains is high. The methods applied were found useful for typing B. henselae isolates, providing tools for epidemiological and clinical follow-up studies.
从德国弗赖堡地区家猫的血培养物中获得了17株汉赛巴尔通体分离株，对其基因异质性进行了检测。基于不同的DNA指纹图谱方法，包括脉冲场凝胶电泳（PFGE）、肠杆菌基因间重复一致序列（ERIC）-PCR、重复外显子回文序列（REP）PCR和任意引物（AP）-PCR，在这些分离株中鉴定出三种不同的变体（变体I至III）。变体I包括6株，变体II包括10株，变体III仅包括1株。通过所有使用的方法，这些分离株可与模式菌株休斯顿-1（指定为变体IV）明显区分开来。先前发表的16S rDNA型特异性扩增区分了汉赛巴尔通体分离株的两种类型（16S rRNA类型1和2）。大多数分离株（17株中的16株），包括所有变体I和II，为16S rRNA类型2。只有一株分离株（变体III）和休斯顿-1菌株（变体IV）属于16S rRNA类型1。对来自三个变体（I至III）中每个变体的一株代表性菌株的16S rDNA序列进行比较，证实了16S rRNA型特异性PCR获得的结果。变体I和变体II的序列相同，而变体III的序列在三个位置不同。本研究中应用的所有方法都能对分离株进行亚型分析。PFGE和ERIC-PCR对汉赛巴尔通体菌株亚型分析具有最高的鉴别潜力，而使用M13引物的AP-PCR在四种变体之间显示出非常明显的区分。我们的结果表明，汉赛巴尔通体菌株的基因异质性很高。所应用的方法被发现可用于对汉赛巴尔通体分离株进行分型，为流行病学和临床随访研究提供工具。
