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用于检测血样中杆菌状巴尔通体的聚合酶链反应方法评估

Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples.

作者信息

Gomes Cláudia, Martinez-Puchol Sandra, Pons Maria J, Bazán Jorge, Tinco Carmen, del Valle Juana, Ruiz Joaquim

机构信息

ISGlobal, Barcelona Ctr. Int. Health Res. (CRESIB), Hospital Clínic - Universitat de Barcelona, Barcelona, Spain.

Faculty of Health Sciences, School of Medicine, Centro de Investigación de la Universidad Peruana de Ciencias Aplicadas (UPC), Lima, Peru.

出版信息

PLoS Negl Trop Dis. 2016 Mar 9;10(3):e0004529. doi: 10.1371/journal.pntd.0004529. eCollection 2016 Mar.

Abstract

BACKGROUND

The lack of an effective diagnostic tool for Carrion's disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches.

METHODOLOGY/PRINCIPAL FINDINGS: We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay.

CONCLUSIONS/SIGNIFICANCE: From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion's disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques.

摘要

背景

缺乏针对卡里翁病的有效诊断工具会导致误诊、错误治疗,以及流行地区无症状携带者的持续存在。传统的聚合酶链反应(PCR)方法已被报道为一种诊断技术。然而,这些技术的检测限尚不清楚,其在低菌血症病例中的实用性也不明确。本研究的目的是评估三种PCR方法的检测限。

方法/主要发现:我们确定了三种不同PCR方法的检测限:巴尔通体特异性16S核糖体RNA、鞭毛蛋白及其基因。我们还评估了干血斑用作样本运输系统的可行性。我们的结果表明,16S rRNA PCR是检测限最低的方法,为5 CFU/μL,因此是所研究的最佳诊断PCR工具。干血斑会降低检测的灵敏度。

结论/意义:在所测试的PCR方法中,16S rRNA PCR方法最适合用于卡里翁病急性病例的直接血液检测。然而,其用于干血斑样本时,虽然便于农村地区运输样本的管理,但灵敏度略有下降。通过PCR检测低菌血症或无症状携带者的存在情况存疑,这表明需要寻找更新的更灵敏的技术。

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