Design, Optimization and Evaluation of a Polymerase Chain Reaction for Detection of Borrelia spp.
Journal Title: Advances in Clinical and Experimental Medicine - Year 2013, Vol 22, Issue 5
Abstract
<strong>Background</strong>. Lyme borreliosis and relapsing fever are important zoonotic diseases worldwide and the improvement of diagnostic strategies is a prioritized task considering the morbidity of these diseases in some areas. PCR based methods appear to be of utmost importance because of the high sensitivity and specificity of these assays.<br /><strong>Objectives. </strong>To obtain a molecular method based on PCR for the detection of the genus <em>Borrelia </em>infection in different specimens.<br /><strong>Results. </strong>Sets of reported primers were evaluated “in silico” and they did not fulfill the proposal parameters. On the other hand, the two new, designed sets of primers were theoretically efficient for <em>Borrelia </em>DNA amplification. PCR procedures with these primers were standardized with borrelial DNA and optimum annealing temperatures, primer concentrations and reaction cycle numbers were determined. The PCR analytical sensitivity was 10 genomes per reaction for each technique. Both PCR were highly specific to different <em>Borrelia </em>species DNA and to samples (sera, cerebrospinal liquids and hard ticks) infected artificially with a <em>Borrelia </em>strain, visualizing the amplification of the expected DNA fragment. No amplification was obtained when other microorganisms were used. 36 human clinical samples were negatives in a preliminary study.<br /><strong>Conclusions. </strong>Both sets of primers with their respective PCR protocols showed similar results, which suggest that each one can be used indistinctly in detecting <em>Borrelia </em>spp., mainly in countries where the situation of these diseases are unknown.
Authors and Affiliations
Angel Noda, Islay Rodríguez, Brian Mondeja, Carman Fernándeze
Keywords
Related Articles
- EP ID EP115385
- DOI -
- Views 93
- Downloads 0
Influence of Central Noradrenergic System Lesion on the Serotoninergic 5-HT3 Receptor Mediated Analgesia in Rats
<strong>Background. </strong>Monoaminergic pathways, impinging an adrenergic and 5-HT3 serotonin receptors, modulate nociceptive transmission, but their mechanisms and interactions has not been clarified yet.<br /><stron...
Przeciwciała przeciwko oksydowanym LDL oraz przeciwko kompleksom oxLDL-β2GPI u chorych na tocznia rumieniowatego układowego
[b]Cel pracy.[/b] Ocena stężenia przeciwciał skierowanych przeciwko cząsteczkom oxLDL (utlenowanych lipoprotein o niskiej gęstości) oraz przeciwko kompleksom zawierających oxLDL i beta2-glikokoproteinę I (oxLDL-β2GPI) w...
Depression in Women Aged 75–89 – Predisposing Factors and Preventive Measures
[b]Background.[/b] There are analyses showing the relationship between low and irregular physical activity and the risk of more frequent occurrences of depression symptoms in the future. There are studies that do not pro...
Quantitative Diagnosis of Early-Stage Liver Cirrhosis with Contrast-Enhanced Ultrasound – A Clinical Study
[b]Objectives.[/b] To investigate the quantitative diagnosis value of contrast-enhanced ultrasound (CEUS) for early-stage liver cirrhosis. [b]Material and Methods.[/b] A total of 15 healthy subjects and 15 patients wit...
The Impact of Rehabilitation Carried out Using the Masgutova Neurosensorimotor Reflex Integration Method in Children with Cerebral Palsy on the Results of Brain Stem Auditory Potential Examinations
[b]Background.[/b] Rehabilitation therapy in children with neuromotor development disorders can be carried out with the use of various methods. [b]Objectives.[/b] The aim of this study was to determine the efficiency o...