Cíl studie: Rozlišit genotypy a subtypy Borrelia burgdorferi sensu lato a Ehrlichia spp. V kulturách, ve vzorcích mozkomíšního moku a krve pacientů a ve tkáních zvířat. Studie byla zaměřena na střední a východní Čechy a Moravu. Materiál: Zkoumáno bylo 16 kmenů borelií, izolovaných z krve a moku pacientů, dále 49 vzorků mozkomíšního moku a krve pacientů s boreliózou, tkáně 80 kusů lovné zvěře a krev 55 krav. Metody: Diferenciaci a kvantifikaci obou patogenů umožnilo užití LightCycler PCR v reálném čase s příměry 16S rRNA, OspA, recA genů a přímá PCR-sekvenační analýza produktů. Výsledky: Prokázali jsme 11 kmenů Borrelia garinii, OspA-typy 5,6,4 a pět kmenů B. afélii. B. burgdorferi sensu stricto nebyla v kulturách prokázána. Sekvenační analýzou PCR produktů ze 49 vzorků mozkomíšního moku jsme prokázali B. burgdorferi s.s. v 20,4 %, B. afzeJii v 12,2 %, B. garinii, subtypy 6, 5, 4 a 3 v 61,3 %, z nichž 8,2 % vzorků obsahovalo kombinace blízkých OspA-typů. Koinfekce s EhrJichia phagocytophiia byla zjištěna v 6,1 % vzorků. E. phagocytophiia subtyp Frankonia I a II jsme prokázali u 12,5 % lovné zvěře a u 9 % skotu. B. burgdorferi sensu strigo a B. garinii byla zjištěna u 17,5 % lovné zvěře. Light Cycler PCR v reálném čase byla méně sensitivní v tekutinách pacientů než v kulturách a v tkáních zvířat. Závěr: Prokázali jsme, že neuroboreliózu mohou působit všechny tři genospecies B. burgdorferi sensu lato. Průběh infekce a symptomy mohou ovlivnit různé OspA- subtypy.
Purpose of the study: To differentiate the genotypes and subtypes of Borreiia burgdorferi sensu lato and Ehrlichia spp. in cultures, in samples of patients' cerebrospinal fluid and blood and in animal tissues. The study focuses on southern and eastern Bohemia and on Moravia. Material: The investigation involved 16 borrelia strains isolated from the blood and CSP of patients and further 49 samples of CSF and blood of borreliosis patients, tissues from 80 pieces of game animals and blood from 55 cows. Methods: The differentiation and quantification of the two pathogenes was done using real-time LightCycler PCR with the primers loS rRNA, OpA, recA genes and direct PCR sequential analysis of the products. Results: We demonstrated 11 strains of Borreiia garinii, OspA types 5, 6, 4 and 5 strains of B. afzelii. B. burgdorferi sensu stricto was not demonstrated in the cultures. The sequential analysis PCR of the products from 49 CSF samples demonstrated B, burgdorferi s.s. in 20.4 %, B. afzelii in 12.2 %, B. garnii subtypes 6, 5, 4 and 3 in 61.3 % - out of these 8.2 % of the samples contained combinations similar to OspA types. Co-infection with Ehrlichia phagocytophila was found in 6.1 % of samples. E. phagocytophila subtype Frankonia I and II was demonstrated in 12.5 % of game animals and in 9 % of cattle. B. burgdorferi sensu stricto and B. garinii were identified in 17.5 % of game. Real-time LightCycler PCR was less sensitive in patients' fluids than in cultures and animal tissues. Conclusions: We demonstrated that all three genotypes of B. burgdorferi sensu lato are capable of causing neuroborreliosis. The course of the infection and its symptoms may be affected by various OspA subtypes.
- MeSH
- Borrelia genetics classification pathogenicity MeSH
- Ehrlichia genetics pathogenicity MeSH
- Ehrlichiosis diagnosis microbiology MeSH
- Research Support as Topic MeSH
- Humans MeSH
- Lyme Disease diagnosis microbiology MeSH
- Reverse Transcriptase Polymerase Chain Reaction methods MeSH
- Signs and Symptoms diagnosis MeSH
- Sequence Analysis methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Review MeSH
- Comparative Study MeSH
Tento přehled se zabývá využitím různých elektrochemických detekčních systémů v průtokovýchmetodách analýzy léčiv (jako jsou metody FIA a SIA). Přehled pokrývá období od roku 1988-1998a zahrnuje 78 odkazů. Stanovované léčivé látky jsou seřazeny podle funkčních skupin podléhajícíchelektrochemické transformaci; analytická data zahrnují detekční podmínky, detekční limity a rozsahy kvantity. Rovněž se zabývá výhodami a nevýhodami amperometrické detekce léčiv v průtokových systémech.
The present review dealing with the use of various electrochemical detection systems in flowmethods of analysis of drugs (such as FIA and SIA techniques). The review covers the period of 1988to 1998 and involves 78 references. The drugs determined are arranged according to the functionalgroups undergoing electrochemical transformation; for all the analytes data on the detectionconditions, detection limits and ranges of quantitation are included. Advantages and drawbacks ofamperometric detection of drugs in flow systems are discussed.
Statistics in practice
2nd rev.ed xii, 314 s.
- MeSH
- Clinical Trials as Topic MeSH
- Publication type
- Clinical Trial MeSH
- Conspectus
- Patologie. Klinická medicína
- NML Fields
- diagnostika
The aim of the present study was to determine the profile of different inflammatory molecules in serum and cerebrospinal fluid (CSF) during invasive meningococcal disease (IMD). Their relationship with IMD severity was also assessed. A cohort of 12 patients with IMD was investigated. Paired serum and CSF samples were obtained at the time of diagnostic and follow-up lumbar puncture and were examined using Luminex analysis. IMD severity correlated with serum interleukin-6 (IL-6) and interleukin-1 receptor antagonist (IL-1 ra) on admission. Furthermore, the CSF levels of IL-1 beta, IL-1 ra, IL-6, IL-8, macrophage inflammatory protein-1 beta (MIP-1 beta), and monocyte chemoattractant protein-1 (MCP-1) were significantly higher than their respective serum levels. The strongest correlations were found between serum concentrations of IL-1 beta and IL-1 ra, IL-6, IL-8, and MIP-1 beta, whereas the strongest correlations in CSF were found between endotoxin and IL-8, IL-17, MIP-1 beta, and MCP-1. As was expected, the concentrations of inflammatory molecules in both serum and CSF significantly decreased after antibiotic treatment. With regard to kinetics, a severe course of IMD correlated positively with rapid declines of CSF IL-6 and cortisol levels. Sequential multiple analyses revealed patterns of inflammatory responses that were associated with the severity of IMD, as well as with the compartmentalization and kinetics of the immune reaction.
- MeSH
- Anti-Bacterial Agents therapeutic use MeSH
- Biomarkers MeSH
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Inflammation Mediators analysis MeSH
- Meningococcal Infections drug therapy microbiology pathology MeSH
- Adolescent MeSH
- Young Adult MeSH
- Cerebrospinal Fluid chemistry MeSH
- Serum chemistry MeSH
- Severity of Illness Index MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
A review is presented on the state of the art of the chemiluminescence analysis of pharmaceuticals by the two most relevant automated controlled-flow methodologies--flow-injection analysis (FIA) and sequential-injection analysis (SIA). The current chemiluminometric applications of FIA and SIA in pharmaceutical analysis are discussed with special emphasis on the analytical figures of merit and sample matrix characteristics. The review involving 211 references and covering papers published between 2001 and 2006 is divided into several sections according to the fundamental types of chemiluminescence systems employed.
