Clarridge J E, Raich T J, Pirwani D, Simon B, Tsai L, Rodriguez-Barradas M C, Regnery R, Zollo A, Jones D C, Rambo C
Laboratory Service, Veterans Affairs Medical Center, Houston, Texas, USA.
J Clin Microbiol. 1995 Aug;33(8):2107-13. doi: 10.1128/jcm.33.8.2107-2113.1995.
We wished to develop a cost-effective, rapid strategy to detect and identify Bartonella species in the clinical laboratory and to determine the prevalence of Bartonella infection in the Houston veteran population. Bartonella colonies were identified by colony morphology, Gram stain, RapID ANA, repetitive extragenic palindromic-PCR (REP-PCR) and whole-cell fatty acid (CFA) analysis, and these methods were compared for their usefulness. A new test order for "Rochalimaea culture" (the genus Bartonella was previously known as the genus Rochalimaea) was instituted, and in addition, all blood specimens submitted for fungal culture (obtained in an isolator tube) were processed for Bartonella culture. Over a 16-month period we isolated Bartonella henselae from only 0.4% (2 of 533) of total cultures but from 1% (2 of 204) of human immunodeficiency virus-positive patients. After sufficient growth, identification of the Bartonella isolates to the species level could be obtained in 2 days. The REP-PCR allowed discrimination of all known species, whereas CFA analysis distinguished all except B. henselae and Bartonella quintana. The RapID ANA results failed to differentiate between B. henselae and B. quintana, and results for other species differed by only one or two tests. Blood obtained from a kitten which had been introduced into the household of one patient 2 months before the onset of fever yielded a Bartonella strain which was shown to be different from the strain from the patient and distinct from other Bartonella species by a combination of REP-PCR, CFA, and growth characteristics. Subsequent analysis of the citrate synthase gene sequence showed only an 86% similarity with any of the other known Bartonella species, suggesting that this isolate represents a distinct, previously uncharacterized species of Bartonella.
我们希望开发一种经济高效、快速的策略,用于在临床实验室中检测和鉴定巴尔通体菌种,并确定休斯顿退伍军人人群中巴尔通体感染的患病率。通过菌落形态、革兰氏染色、RapID ANA、重复外显子回文PCR(REP-PCR)和全细胞脂肪酸(CFA)分析来鉴定巴尔通体菌落,并比较这些方法的实用性。制定了一项新的“罗卡利马氏体培养”检测项目(巴尔通体属以前被称为罗卡利马氏体属),此外,所有提交进行真菌培养的血液标本(在隔离管中采集)都进行了巴尔通体培养处理。在16个月的时间里,我们仅从总培养物的0.4%(533份中的2份)中分离出了汉赛巴尔通体,但从1%(204名人类免疫缺陷病毒阳性患者中的2名)的患者中分离出了该菌。在充分生长后,2天内即可将巴尔通体分离株鉴定到种水平。REP-PCR能够区分所有已知菌种,而CFA分析除了汉赛巴尔通体和五日热巴尔通体之外,能区分所有其他菌种。RapID ANA结果无法区分汉赛巴尔通体和五日热巴尔通体,其他菌种的结果仅相差一两项检测。从一名患者发热前2个月引入其家中的一只小猫身上采集的血液,培养出了一株巴尔通体菌株,通过REP-PCR、CFA和生长特性的综合分析表明,该菌株与患者的菌株不同,也与其他巴尔通体菌种不同。随后对柠檬酸合酶基因序列的分析表明,它与任何其他已知巴尔通体菌种的相似性仅为86%,这表明该分离株代表了一种独特的、以前未被鉴定的巴尔通体菌种。