SANTRAUKA Serologinių sifilio ir Laimo boreliozės reakcijų kryžminis reaktyvumas Darbo autorė: Tatjana Subočiūtė Darbo vadovas: med. m. dr. Andrius Vagoras Darbo tikslas: nustatyti VULSK diagnostikos centre rutiniškai naudojamų serologinių sifilio ir Laimo boreliozės testų galimai klaidingai teigiamų rezultatų dažnumą dėl kryžminio viena iš šių infekcijų užsikrėtusiųjų pacientų kraujo serumų reaktyvumo. Darbo metodai: Tyrimai buvo atlikti Vilniaus universiteto ligoninės Santariškių klinikos (VULSK) Laboratorinės diagnostikos centro Mikrobiologijos laboratorijos Infekcijų žymenų tyrimų padalinyje. Serumai buvo surinkti ir ištirti nuo 2005 m. vasario iki 2006 m. kovo mėn. Tyrimo medžiagą sudarė 40 serumų. 23 serumai dėl sifilio tyrimų buvo gauti iš VULSK Dermatovenerologijos centro. 8 serumai dėl Laimo ligos tyrimų buvo gauti iš VULSK Dermatovenerologijos centro. UAB „Endemik” atrinkti 9 serumai ištirti dėl Laimo ligos. Visi į tyrimą įtraukti serumai buvo ištirti šiomis reakcijomis: 1) RPR su titru, 2) TPHA, 3) Treponema pallidum ELISA, 4) Treponema pallidum imunofluorescencija, 5) Borrelia burgdorferi imunofluorescencija. Tyrimo rezultatai: Ištyrus penkiomis reakcijomis serumus (RPR, TPHA, Treponema pallidum IFR, Treponema pallidum IFA bei Borrelia burgdorferi IFR), buvo pastebėtas kryžminis serologinis reaktyvumas tarp Borrelia burgdorferi IFR ir Treponema pallidum IFR reakcijų. Mūsų darbo duomenimis, tiriant 17 serumų (teigiamus dėl Laimo ligos) kryžminis reaktyvumas... [toliau žr. visą tekstą] / SUMMARY Cross - reactivity in serological test for Lyme disease and syphilis Author: Tatjana Subočiūtė Supervisor: Andrius Vagoras, MD Work purpose: to determine the incidence of possibly false positive results of serological syphilis and Lyme borreliosis tests, routinely used in VULSK Diagnostics centre, occurring because of cross reactivity of patients, having one of these disease, blood. Work methods: the research was carried out in Vilnius University Hospital Santariškių Klinikos (VULSK) Infection Markers Research Department of Microbiological Laboratory in Laboratory Diagnostics Centre. Serum samples were collected and tested from February, 2005, till March, 2006. The investigation material was composed of 40 serum samples. 23 serum samples for syphilis test came from VULSK Centre of Dermatovenereology. 8 serum samples for Lyme borreliosis test were received from VULSK Centre of Dermatovenereology. 9 serum samples for Lyme borreliosis test were picked by UAB “Endemik”. All the samples, included into the research, were tested by following reactions: 1) RPR with titer, 2) TPHA, 3) Treponema pallidum ELISA, 4) Treponema pallidum immunofluorescence, 5) Borrelia burgdorferi immunofluorescence. Research results: After testing the serums by five reactions (PRP, TPHA, Treponema pallidum FTA – ABS, Treponema pallidum ELISA and Borrelia burgdorferi immunofluorescence), a serological cross reactivity between Borrelia burgdorferi immunofluorescence and Treponema pallidum FTA - ABS... [to full text]

Diss. (sammanfattning) Linköping : Univ., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2006. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. "May 2007" Includes bibliographical references.

Thesis (M.S.)--Michigan State University. Dept. of Fisheries and Wildlife, 2008. / Title from PDF t.p. (viewed on July 30, 2009) Includes bibliographical references (p. 176-183). Also issued in print.

Thesis (Ph. D.) -- University of Texas Southwestern Medical Center at Dallas, 2005. / Vita. Bibliography: 284-323.

The aim of the study was to analyze the infectious potential of the newly described species, B. americana and B. carolinensis, studied on the laboratory model mammals mice. Our goal was to analyze and compare the vectorial capacity of two different tick vectors, Amblyomma americanum and Ixodes ricinus, in acquiring and transmition of both spirochete species to the host. The results of this study confirmed that ticks A. americanum and I. ricinus are capable to maintain and transmit B. americana and B.carolinensis.We confirmed that both analysed spirochete species, B. carolinensis and B. americana, showed the potential to develop the disease in laboratory model mammal, which indirectly support the fact that both spirochete species might be concidered as the risk factors in the area where they are distributed. Our results shows that A. americanum is able to transmit both spirochete species, which increases that risk of acquiring the Lyme disease to human population in the area of distribution of A. americanum

abstract: Glycosaminoglycans (GAGs) are a class of complex biomolecules comprised of linear, sulfated polysaccharides whose presence on cell surfaces and in the extracellular matrix involve them in many physiological phenomena as well as in interactions with pathogenic microbes. Decorin binding protein A (DBPA), a Borrelia surface lipoprotein involved in the infectivity of Lyme disease, is responsible for binding GAGs found on decorin, a small proteoglycan present in the extracellular matrix. Different DBPA strains have notable sequence heterogeneity that results in varying levels of GAG-binding affinity. In this dissertation, the structures and GAG-binding mechanisms for three strains of DBPA (B31 and N40 DBPAs from B. burgdorferi and PBr DBPA from B. garinii) are studied to determine why each strain has a different affinity for GAGs. These three strains have similar topologies consisting of five α-helices held together by a hydrophobic core as well as two long flexible segments: a linker between helices one and two and a C-terminal tail. This structural arrangement facilitates the formation of a basic pocket below the flexible linker which is the primary GAG-binding epitope. However, this GAG-binding site can be occluded by the flexible linker, which makes the linker a negative regulator of GAG-binding. ITC and NMR titrations provide KD values that show PBr DBPA binds GAGs with higher affinity than B31 and N40 DBPAs, while N40 binds with the lowest affinity of the three. Work in this thesis demonstrates that much of the discrepancies seen in GAG affinities of the three DBPAs can be explained by the amino acid composition and conformation of the linker. Mutagenesis studies show that B31 DBPA overcomes the pocket obstruction with the BXBB motif in its linker while PBr DBPA has a retracted linker that exposes the basic pocket as well as a secondary GAG-binding site. N40 DBPA, however, does not have any evolutionary modifications to its structure to enhance GAG binding which explains its lower affinity for GAGs. GMSA and ELISA assays, along with NMR PRE experiments, confirm that structural changes in the linker do affect GAG-binding and, as a result, the linker is responsible for regulating GAG affinity. / Dissertation/Thesis / Doctoral Dissertation Biochemistry 2015

The aim of present master’s thesis was to compare the results of serological methods for diagnosing borreliosis that are commonly used in Spadia laboratories (ELISA, immunoblots) in terms of recommendation on how and when to indicate and interpret them. The theoretical part is focusing on the characteristics and history of borreliosis, microbiological description of Borrelia, immune system and pathogenesis of the disease as well as the therapy and prevention. The experimental part is focusing on the analysis of results obtained from common examinations of antibodies to Borrelia made in Spadia Lab laboratories from January 1st 2014 to December 31st 2015. Screening of antibodies to Borrelia made by ELISA in IgM and IgG was done for all samples according to recommendation of CDC. In 2014 the ELISA screening was done using ELISA kits from Euroimmun and Evolis sample processors whereas in 2015 it was done using DiaSorin’s CLIA kits on Liaison analyser. Positive results were then confirmed by Westernblot or lineblot alternatively if the physician did not ask otherwise. It must be remembered that ELISA and Westernblot belong among serological methods that are using antibodies, i.e. substances produced by the immune system. The immune system plays the key role in protecting the body against infection and the antibodies are its important tool. Serological methods belong among immunoassay methods, which is still not standardized. Diagnosis of infections cann‘t be based only on antibody testing. It is necessary to assess the results in the context of the entire clinical picture, history and in the case of antibodies it is recommended retesting with an interval.

Although Lyme disease is the most frequently reported vector-borne illness in the United States, recent evidence from the CDC suggests that Lyme disease incidence in the United States may be much higher than reported. Lyme disease symptoms can be mistaken for a wide variety of diseases, which can complicate the diagnosis. To date, no diagnostic criteria analysis has been conducted examining the association between sociodemographic variables (sex and age) and seasonality of infection with the severity and symptomology found in Lyme disease cases. Using the CDC's outbreak investigation model, a primary case/control study was conducted using the ROSS Scale to collect data. Comparisons were made between a Lyme disease-diagnosed group (n = 203) and a convenience sample of non-Lyme disease patients (n = 388). Novel symptom patterns were found to significantly predict a diagnosis of Lyme disease. Odds ratio results revealed a positive association between musculoskeletal (OR = 11; 95% CI), neurological (OR = 12; 95% CI), cognitive (OR = 10; 95% CI), and cutaneous (OR = 144; 95% CI) symptoms frequency and severity and the diagnosis of Lyme disease. In addition, overall symptom frequency and severity scores displayed significant differences between cases and controls, between males and females, and among certain age groups. No correlation was found between symptom frequency and severity with the seasonality of infection. Current diagnostic tools search for antibodies to the Borrelia bacteria, but antibody production takes a few weeks. The results of this study help identify at-risk patients based on the presentation and severity of Lyme disease symptoms when antibodies are not present in measureable quantities in the blood stream, allowing for earlier diagnosis.