Rapid identification and differentiation of Bartonella species using a single-step PCR assay
Rapid identification and differentiation of Bartonella species using a single-step PCR assay
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DOI：
10.1128/jcm.38.5.1717-1722.2000
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发表时间：
2000-05-01
影响因子：
9.4
通讯作者：
Breitschwerdt, EB
中科院分区：
文献类型：
--
作者：
Jensen, WA;Fall, MZ;Breitschwerdt, EB
据报道，五种巴尔通体感染人类，并导致各种难以诊断的疾病。据报道，四种巴尔通体会感染猫和狗，其中两种被认为是人畜共患病原体。巴尔通体属感染的诊断受到巴尔通体属物种缓慢、挑剔的生长特征的阻碍。我们报告的发展，一个单步PCR为基础的检测和医学相关的巴尔通体物种的分化。PCR介导的16 S-23 S rRNA基因间区扩增导致每个巴尔通体属物种的产物具有独特的大小，从而允许区分而不需要限制性片段长度多态性分析或扩增产物的测序。使用已知感染汉赛巴尔通体（B）的动物的特征分离株和血液样本，确定单步PCR检测区分巴尔通体种属的能力。clarridgeiae或B.温氏亚种berkhoffi。用来自B的血液样品测定了单步PCR检测相对于体外培养的灵敏度。感染了汉赛拉菌的猫B。汉赛氏菌靶DNA从100%的大于50 CFU/ml的样品和80%的10 - 30 CFU/ml的样品中扩增。报告中描述的一步测定加快了医学相关巴尔通体属种的基于PCR的检测和区分。
Five species of Bartonella have been reported to infect humans and cause a variety of diseases that can be difficult to diagnose. Four species of Bartonella have been reported to infect cats and dogs, and two of these species are considered to be zoonotic pathogens. Diagnosis of Bartonella infections is hampered by the slow, fastidious growth characteristics of Bartonella species. We report on the development of a single-step PCR-based assay for the detection and differentiation of medically relevant Bartonella species. PCR-mediated amplification of the 16S-23S rRNA intergenic region resulted in a product of a unique size for each Bartonella species, thereby allowing differentiation without the necessity of restriction fragment length polymorphism analysis or sequencing of the amplified product. The ability of the single-step PCR assay to differentiate between Bartonella species was determined with characterized isolates and blood samples from animals known to be infected with either Bartonella henselae, B. clarridgeiae, or B. vinsonii subsp. berkhoffi. The sensitivity of the single-step PCR assay relative to that of in vitro culture was determined with blood samples from B. henselae-infected cats. B. henselae target DNA was amplified from 100% of samples with greater than 50 CFU/ml and 80% of samples with 10 to 30 CFU/ml. The single-step assay described in the report expedites PCR-based detection and differentiation of medically relevant Bartonella species.