We investigated the presence of glycoproteins in Borrelia burgdorferi. We did not find any evidence for glycosylation of the major outer membrane proteins OspA and OspB or the structural flagellar proteins FlaB and FlaA. We suggest that glycoproteins present on the surface of B. burgdorferi may be tightly bound culture medium glycoproteins.

• MeSH
• Antigens, Bacterial genetics   metabolism   MeSH
• Antigens, Surface genetics   metabolism   MeSH
• Bacterial Vaccines genetics   metabolism   MeSH
• Borrelia burgdorferi genetics   metabolism   ultrastructure   MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Cryoelectron Microscopy MeSH
• Flagellin genetics   metabolism   MeSH
• Glycosylation MeSH
• Spectrometry, Mass, Electrospray Ionization MeSH
• Lipoproteins genetics   metabolism   MeSH
• Molecular Sequence Data MeSH
• Bacterial Outer Membrane Proteins genetics   metabolism   MeSH
• Amino Acid Sequence MeSH
• Sequence Homology, Amino Acid MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• Antigens, Surface MeSH
• Bacterial Vaccines MeSH
• flaA protein, bacteria MeSH Browser
• flaB flagellin MeSH Browser
• Flagellin MeSH
• Lipoproteins MeSH
• OspA protein MeSH Browser
• OspB protein, Borrelia burgdorferi MeSH Browser
• Bacterial Outer Membrane Proteins MeSH

A simple assay by polymerase chain reaction was used for the of detection of Borrelia burgdorferi, causative agent of Lyme borreliosis (LB). It involves no DNA purification and is based on the amplification of a specific region of ospA gene of B. burgdorferi, followed by direct detection of the PCR product with SYBR Green I by agarose gel electrophoresis. The method was used to analyze samples from patients with LB diagnosis, with presumable infection with the LB spirochete, those with unclear clinical symptoms and after the course of an antibiotic treatment. Spirochetal DNA was detected by PCR even in contaminated samples in which B. burgdorferi was overgrown by fungi and other bacteria. Spirochetal DNA was detected and borrelia species was identified in cerebrospinal fluid of two patients hospitalized with the diagnosis "fever of unknown origin". Western blot and ELISA were negative in both cases. Total analysis of 94 samples from the hospital in Ceské Budejovice (South Bohemia, Czechia) showed infection with B. burgdorferi sensu stricto in 11% and B. garinii in 15% of cases. The highest prevalence was found for B. afzelii (43%). Co-infection was confirmed in 24 % of the analyzed symplex; 7% of samples that were B. burgdorferi sensu lato positive gave no results in DNA amplification with B. burgdorferi sensu stricto-, B. garinii- and B. afzelii-specific primers. The proposed reliable, rapid, unexpensive and specific technique could form the basis of laboratory tests for routine detection and identification of Lyme-disease spirochete in different samples.

• MeSH
• Antigens, Surface genetics   MeSH
• Bacterial Vaccines MeSH
• Benzothiazoles MeSH
• Borrelia burgdorferi Group classification   genetics   immunology   isolation & purification   MeSH
• Quinolines MeSH
• Diamines MeSH
• DNA, Bacterial analysis   isolation & purification   MeSH
• Electrophoresis, Agar Gel MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Genotype MeSH
• Immunoglobulin G blood   MeSH
• Immunoglobulin M blood   MeSH
• Humans MeSH
• Lipoproteins genetics   MeSH
• Lyme Disease diagnosis   MeSH
• Cerebrospinal Fluid microbiology   MeSH
• Organic Chemicals metabolism   MeSH
• Polymerase Chain Reaction methods   MeSH
• Bacterial Outer Membrane Proteins genetics   MeSH
• Antibodies, Bacterial blood   MeSH
• Sensitivity and Specificity MeSH
• Serum microbiology   MeSH
• Bacterial Typing Techniques MeSH
• Blotting, Western MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, Surface MeSH
• Bacterial Vaccines MeSH
• Benzothiazoles MeSH
• Quinolines MeSH
• Diamines MeSH
• DNA, Bacterial MeSH
• Immunoglobulin G MeSH
• Immunoglobulin M MeSH
• Lipoproteins MeSH
• Organic Chemicals MeSH
• OspA protein MeSH Browser
• Bacterial Outer Membrane Proteins MeSH
• Antibodies, Bacterial MeSH
• SYBR Green I MeSH Browser

Borrelial glycoconjugates were localized by labeled lectins on ultrathin cryosections and on surfaces of intact negatively stained bacteria. Protein-saccharide complexes in these glycoconjugates were partially characterized by means of enzyme deglycosylation and mild alkali pretreatment of cryosections. The results of labeling were examined by transmission electron microscopy. Statistically evaluated results (relative labeling index, chi2 test) of gold labeling indicated that surfaces of Borrelia burgdorferi strain B31 and external (outer) membrane vesicles (MVs) were covered with glycoconjugates containing O-glycosidically linked N-acetyl-D-galactosamine (GalNAc) and N-glycosidically linked N-acetyl-D-glucosamine (GlcNAc). The presence of N-linked GalNAc, sialic acid, mannose and fucose on the surfaces of outer membranes and MVs was probably due to an adherence of BSK-H medium components, especially rabbit serum, to Borrelia surfaces.

• MeSH
• Borrelia burgdorferi metabolism   pathogenicity   ultrastructure   MeSH
• Cell Membrane metabolism   ultrastructure   MeSH
• Cryoelectron Microscopy MeSH
• Glycoconjugates chemistry   metabolism   MeSH
• Rabbits MeSH
• Lectins metabolism   MeSH
• Humans MeSH
• Lyme Disease microbiology   MeSH
• In Vitro Techniques MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Glycoconjugates MeSH
• Lectins MeSH

Ixodes ricinus ticks were collected by random collections from western and central Slovakia during the years 1996-98. The midgut content of 240 ticks was examined by dark-field microscopy and submitted for cultivation for the presence of borrelias. Spirochetes were found in 21 unfed and host-seeking adults and nymphs (8.8%). By the analysis of restriction fragment length polymorphism (RFLP) one sample from unfed I. ricinus male from western Slovakia was identified as a triple infection of Borrelia burgdorferi sensu stricto, B. garinii and B. afzelii. The simultaneous presence of different B. burgdorferi genospecies in one midgut sample (triple infection in the tick) could be observed only after the multipart amplification of denaturated DNA and subsequent pooling of the products for further analysis.

• MeSH
• Borrelia burgdorferi classification   genetics   isolation & purification   MeSH
• Genotype MeSH
• Ixodes microbiology   MeSH
• Polymerase Chain Reaction MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH