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Multi-Locus Sequence Typing of Bartonella bacilliformis DNA Performed Directly from Blood of Patients with Oroya's Fever During a Peruvian Outbreak.Pons, Maria J, Silva-Caso, Wilmer, Del Valle Mendoza, Juana, Ruiz, Joaquim 01 February 2016 (has links)
Background
Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected tropical
poverty-linked illness. This infection is endemic of Andean regions and it is estimated that
approximately 1.7 million of South Americans are at risk. This bacterium is a fastidious slow
growing microorganism, which is difficult and cumbersome to isolate from clinical sources,
thereby hindering the availability of phylogenetic relationship of clinical samples. The aim of
this study was to perform Multi Locus Sequence Typing of B. bacilliformis directly in blood
from patients diagnosed with Oroya fever during an outbreak in Northern Peru.
Methodology/Principal Findings
DNA extracted among blood samples from patients diagnosed with Oroya’s fever were analyzed
with MLST, with the amplification of 7 genetic loci (ftsZ, flaA, ribC, rnpB, rpoB, bvrR
and groEL) and a phylogenetic analysis of the different Sequence Types (ST) was performed.
A total of 4 different ST were identified. The most frequently found was ST1 present
in 66% of samples. Additionally, two samples presented a new allelic profile, belonging to
new STs (ST 9 and ST 10), which were closely related to ST1.
Conclusions/Significance
The present data demonstrate that B. bacilliformis MLST studies may be possible directly
from blood samples, being a promising approach for epidemiological studies. During the
outbreak the STs of B. bacilliformis were found to be heterogeneous, albeit closely related,
probably reflecting the evolution from a common ancestor colonizing the area. Additional
studies including new samples and areas are needed, in order to obtain better knowledge of
phylogenetic scenario B. bacilliformis
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Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood SamplesGomes, Cláudia, Martinez Puchol, Sandra, Pons, Maria J., Bazán, Jorge, Tinco, Carmen, Del Valle Mendoza, Juana Mercedes, Ruiz, Joaquim 09 March 2016 (has links)
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.
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.
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