ISOLATION AND CHARACTERIZATION BY IMMUNOFLUORESCENCE, SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS, WESTERN-BLOT, RESTRICTION FRAGMENT LENGTH POLYMORPHISM-PCR, 16S RIBOSOMAL-RNA GENE SEQUENCING, AND PULSED-FIELD GEL-ELECTROPHORESIS OF ROCHALIMAEA-QUINTANA FROM A PATIENT WITH BACILLARY ANGIOMATOSIS
ISOLATION AND CHARACTERIZATION BY IMMUNOFLUORESCENCE, SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS, WESTERN-BLOT, RESTRICTION FRAGMENT LENGTH POLYMORPHISM-PCR, 16S RIBOSOMAL-RNA GENE SEQUENCING, AND PULSED-FIELD GEL-ELECTROPHORESIS OF ROCHALIMAEA-QUINTANA FROM A PATIENT WITH BACILLARY ANGIOMATOSIS
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DOI：
10.1128/jcm.32.5.1166-1171.1994
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发表时间：
1994-05-01
影响因子：
9.4
通讯作者：
RAOULT, D
中科院分区：
文献类型：
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作者：
MAURIN, M;ROUX, V;RAOULT, D
从法国人免疫缺陷病毒感染的细菌性血管瘤病患者血液中分离得到一株金雀花，该分离株具有典型的金雀花属植物生长特征，采用典型生化检测时呈惰性。采用免疫荧光、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、免疫印迹（免疫印迹）、柠檬酸合成酶基因限制性片段长度多态性pcr、16S rRNA基因测序和脉冲场凝胶电泳等方法对该分离株进行鉴定，并与参考菌株quintana、Rochalimaea vinsonii和Rochalimaea henselae进行比较。SDS-PAGE、Western blot、TaqI酶限制性内切片段长度多态性pcr和16S rRNA基因测序可以区分3种Rochalimaea，并鉴定法国分离物为R. quintana。然而，用特异性小鼠抗血清的免疫荧光法更容易在种水平上对分离的Rochalimaea进行鉴定。脉冲场凝胶电泳可以区分法国金黄菌和金黄菌，并可作为流行病学工具。
Rochalimaea quintana was isolated from the blood of a French human immunodeficiency virus-infected patient with bacillary angiomatosis, The isolate showed the typical growth characteristics of Rochalimaea species and was inert when typical biochemical testing was used. The purpose of the present work was to characterize and compare this new isolate with reference strains of R. quintana, Rochalimaea vinsonii, and Rochalimaea henselae by using immunofluorescence, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot (immunoblot), restriction fragment length polymorphism-PCR of the citrate synthase gene, 16S rRNA gene sequencing, and pulsed-field gel electrophoresis. SDS-PAGE, Western blot, restriction fragment length polymorphism-PCR with TaqI enzyme, and 16S rRNA gene sequencing could differentiate the three Rochalimaea species and allowed characterization of the French isolate as R. quintana. However, identification of the Rochalimaea isolate to the species level was more easily obtained by immunofluorescence with specific murine antisera. Pulsed-field gel electrophoresis allowed differentiation of the French R. quintana isolate from R. quintana Fuller and may sen e as an epidemiological tool.