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

Abstract

BackgroundThe 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 FindingsWe 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/SignificanceFrom 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.

Molecular Detection of Bartonella bacilliformis in Lutzomyia maranonensis in Cajamarca, Peru: A New Potential Vector of Carrion’s Disease in Peru?

“…The conventional PCR assay used in this study was designed to detect Bartonella genus DNA but is also able to amplify phylogenetically related microorganisms such as Brucella spp. 22 The conventional PCR detection of Mesorhizobium spp., nitrogen-fixing bacteria belonging to the order Rhizobiales, in L. maranonensis is not surprising. These microorganisms are closely related to Bartonella spp.…”

Succinyl-CoA Synthetase: New Antigen Candidate of Bartonella bacilliformis

“…Thus, del Valle et al [ 22 ] detected 21 acute Carrion’s disease cases analyzing 113 previously diagnosed as negative patients using a Bartonella -specific 16S rRNA based PCR. Nonetheless, further studies showed the potential limitation of this technique to detect asymptomatic carriers [ 23 ]. The use of RT-PCR has also been studied [ 16 ], 14 positive acute being detected cases among 63 previously negative children.…”

Genotype patterns for mutations of the cystic fibrosis transmembrane conductance regulator gene: a retrospective descriptive study from Saudi Arabia

“…The PCR products were analyzed by sequencing in both the forward and reverse directions were applied according to currently recommended methods. [8][9][10] Variant detection was scored using publicly available variant databases for CF such as the Cystic Fibrosis Mutation Database (http://www.genet.sickkids.on.ca/ CFTR /Home.html) and The Human Gene Mutation Database -Professional Edition. (http://www.hgmd.…”