INTER-SPECIES AND INTRASPECIES IDENTIFICATION OF BARTONELLA (ROCHALIMAEA) SPECIES
INTER-SPECIES AND INTRASPECIES IDENTIFICATION OF BARTONELLA (ROCHALIMAEA) SPECIES
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DOI：
10.1128/jcm.33.6.1573-1579.1995
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发表时间：
1995-06-01
影响因子：
9.4
通讯作者：
RAOULT, D
中科院分区：
文献类型：
--
作者：
ROUX, V;RAOULT, D
Rochalimaea属的物种，最近更名为Bartonella，越来越受到医学界的关注。五日巴尔通体被报告为战壕热、心内膜炎和杆菌性血管瘤病的病因。B。汉赛拉菌与人类免疫缺陷病毒感染患者的症状和感染如发热、心内膜炎和杆菌性血管瘤病有关，并且与猫抓病的病因有关。如此广泛的感染使得有必要获得种内鉴定工具，以便进行流行病学研究。B，vinsonii，B. elizabethae，B. quintana和B的4个分离物。henselae进行了研究，脉冲场凝胶电泳（PFGE）限制性内切酶NotI，SmaI，和SmaI。获得了四种巴尔通体属的特异性图谱。同一物种的分离物的基因组指纹比较显示DNA限制性内切酶图谱的多态性，并且每个分离物获得了特定的图谱。对B.通过Dice系数、UPGMA（unweighted pair-group method of arithmetic averages）和PackagePhilip程序设计，获得了五加属分离物。在16和23 S rRNA基因之间的基因间间隔区（ITS）上进行PCR扩增，随后使用自动激光荧光DNA测序仪（Pharmacia）进行测序。发现每个B. henselae分离物具有特异性序列，而B.昆塔纳分离物仅分为两组。当对ITS PCR产物进行内切酶限制性分析时，TaqI、HindIII和HaeIII这三种酶允许巴尔通体属的物种鉴定。PCR扩增16 S-23 SrRNA基因ITS后的限制性片段长度多态性可用于菌种的快速鉴定，脉冲场凝胶电泳可作为菌种鉴定的有效方法。
Species of the genus Rochalimaea, recently renamed Bartonella, are of a growing medical interest. Bartonella quintana was reported as the cause of trench fever, endocarditis, and bacillary angiomatosis. B. henselae has been implicated in symptoms and infections of human immunodeficiency virus-infected patients, such as fever, endocarditis, and bacillary angiomatosis, and is involved in the etiology of cat scratch disease. Such a wide spectrum of infections makes it necessary to obtain an intraspecies identification tool in order to perform epidemiological studies. B, vinsonii, B. elizabethae, seven isolates of B. quintana, and four isolates of B. henselae were studied by pulsed-field gel electrophoresis (PFGE) after restriction with the infrequently cutting endonucleases NotI, EagI, and SmaI. Specific profiles were obtained for each of the four Bartonella species. Comparison of genomic fingerprints of isolates of the same species showed polymorphism in DNA restriction patterns, and a specific profile was obtained for each isolate. A phylogenetic analysis of the B. quintana isolates was obtained by using the Dice coefficient, UPGMA (unweighted pair-group method of arithmetic averages), and Package Philip programming. Amplification by PCR and subsequent sequencing using an automated laser fluorescent DNA sequencer (Pharmacia) was performed on the intergenic spacer region (ITS) between the 16 and 23S rRNA genes. It was found that each B. henselae isolate had a specific sequence, while the B. quintana isolates fell into only two groups. When endonuclease restriction analysis of the ITS PCR product was done, three enzymes, TaqI, HindIII, and HaeIII, allowed species identification of Bartonella spp. Restriction fragment length polymorphism after PCR amplification of the 16S-23S rRNA gene ITS may be useful for rapid species identification, and PFGE could be an efficient method for isolate identification.