Norman A F, Regnery R, Jameson P, Greene C, Krause D C
Department of Microbiology, University of Georgia, Athens, USA.
J Clin Microbiol. 1995 Jul;33(7):1797-803. doi: 10.1128/jcm.33.7.1797-1803.1995.
The citrate synthase gene (gltA) of Bartonella henselae was cloned and sequenced to compare genetic divergence among alpha and gamma branches of the class Proteobacteria and to develop enhanced genotypic reagents for B. henselae identification. B. henselae gltA is 1,293 nucleotides in length and 63 to 66% homologous with corresponding gene sequences of Rickettsia prowazekii, Escherichia coli, and Coxiella burnetii. The observed genetic variability suggests that gltA sequences can provide a useful means for studying moderate divergence among related bacteria. Oligonucleotides specific for B. henselae gltA were evaluated for the ability to prime PCR amplification within the alpha and gamma branches of the proteobacteria. Under the conditions used, only B. henselae, Bartonella quintana, and R. prowazekii template DNAs yielded amplification products (approximately 380 bp). DNAs from 28 Bartonella-like isolates of feline origin were amplified by B. henselae primers and analyzed for restriction fragment length polymorphism. The resulting patterns for all 28 isolates were similar or identical to that of the recognized B. henselae strain. Current studies are aimed at optimization of PCR conditions for specificity and sensitivity of amplification of Bartonella sequences from clinical isolates.
克隆并测序了汉赛巴尔通体的柠檬酸合酶基因（gltA），以比较变形菌门α和γ分支之间的遗传差异，并开发用于汉赛巴尔通体鉴定的增强型基因分型试剂。汉赛巴尔通体gltA长度为1293个核苷酸，与普氏立克次体、大肠杆菌和贝氏柯克斯体的相应基因序列具有63%至66%的同源性。观察到的遗传变异性表明，gltA序列可为研究相关细菌之间的适度差异提供有用的手段。评估了针对汉赛巴尔通体gltA的寡核苷酸在变形菌门α和γ分支内引发PCR扩增的能力。在所使用的条件下，只有汉赛巴尔通体、五日热巴尔通体和普氏立克次体的模板DNA产生了扩增产物（约380 bp）。用汉赛巴尔通体引物扩增了来自28株猫源巴尔通体样分离株的DNA，并分析了限制性片段长度多态性。所有28株分离株的结果模式与公认的汉赛巴尔通体菌株相似或相同。目前的研究旨在优化PCR条件，以提高从临床分离株中扩增巴尔通体序列的特异性和敏感性。
