A combined approach for the enhanced detection and isolation of Bartonella species in dog blood samples:: Pre-enrichment liquid culture followed by PCR and subculture onto agar plates
A combined approach for the enhanced detection and isolation of Bartonella species in dog blood samples:: Pre-enrichment liquid culture followed by PCR and subculture onto agar plates
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DOI：
10.1016/j.mimet.2007.01.010
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发表时间：
2007-05-01
影响因子：
2.2
通讯作者：
Breitschwerdt, Edward B.
中科院分区：
文献类型：
--
作者：
Duncan, Ashlee W.;Maggi, Ricardo G.;Breitschwerdt, Edward B.
历史上，直接平板接种、裂解离心或冻融方法已被证明是确认犬巴尔通体属感染的高度不敏感方法。一项前瞻性研究旨在比较使用提交给北卡罗来纳州州立大学病媒传播疾病诊断实验室的样本检测巴尔通体的诊断方法。方法包括间接免疫荧光法，PCR，直接接种血琼脂平板（胰蛋白酶大豆琼脂与5%兔血），并接种到一种新的预富集液体培养基，Bartonello/α-变形菌生长培养基（BAPGM）。连续的研究工作导致了一种组合方法的发展，该方法包括在BAPGM中预富集巴尔通体属的培养物，将液体培养物接种到琼脂平板上，然后使用PCR进行DNA扩增。与直接接种血琼脂平板相比，多方面的方法导致巴尔通体的微生物检测和分离的实质性改善。重要的是，这种方法有助于检测和随后分离自然感染犬血液中两种巴尔通体的单一和合并感染。使用预增菌培养和PCR的组合方法可能有助于诊断犬和其他动物的巴尔通体病。(C)2007 Elsevier B. V.保留所有权利。
Historically, direct plating, lysis centrifugation, or freeze-thaw approaches have proven to be highly insensitive methods for confirming Bartonella species infection in dogs. A prospective study was designed to compare diagnostic methods for the detection of Bartonella using samples submitted to the Vector-Borne Disease Diagnostic Laboratory at North Carolina State University. Methods included indirect immunofluorescence assay, PCR, direct inoculation of a blood agar plate (trypticase soy agar with 5% rabbit blood), and inoculation into a novel preenrichment liquid medium, Bartonello/alpha-Proteobacteria growth medium (BAPGM). Sequential research efforts resulted in the development of a combinational approach consisting of pre-enrichment culture of Bartonella species in BAPGM, sub-inoculation of the liquid culture onto agar plates, followed by DNA amplification using PCR. The multi-faceted approach resulted in substantial improvement in the microbiological detection and isolation of Bartonella when compared to direct inoculation of a blood agar plate, Importantly, this approach facilitated the detection and subsequent isolation of both single and co-infections with two Bartonella species in the blood of naturally infected dogs. The use of a combinational approach of pre-enrichment Culture and PCR may assist in the diagnostic confirmation of bartonellosis in dogs and other animals. (C) 2007 Elsevier B.V. All rights reserved.