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PubMed1998

Peptide-based OspC enzyme-linked immunosorbent assay for serodiagnosis of Lyme borreliosis.

Sera from 210 patients with Lyme borreliosis (LB) were studied by an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (pepC10) comprising the C-terminal 10-amino-acid residues of OspC of Borrelia burgdorferi. We found that 36.3 and 45.0% of the serum samples from patients with erythema migrans (EM) and neuroborreliosis (NB), respectively, displayed immunoglobulin M (IgM) anti-pepC10 reactivities, while these samples rarely (</=8%) displayed IgG antibody reactivities. Sera from patients with acrodermatitis chronica atrophicans did not contain anti-pepC10 antibodies. The diagnostic performance of this newly developed peptide ELISA was compared with those of an ELISA based on the full-length recombinant OspC protein (rOspC) and a commercially available ELISA based on the B. burgdorferi flagellum (Fla). The sensitivity of the IgM pepC10 ELISA was slightly lower (P < 0.04) than that of the rOspC ELISA for EM patients (36.3 versus 43.8%), while there was no difference for NB patients (45.0 versus 48.0%). However, the optical density values obtained by the pepC10 ELISA were generally higher than those obtained by the rOspC ELISA, leading to a significantly better quantitative discrimination between seropositive patients with NB and controls (P < 0.008). The specificity of the pepC10 ELISA was similar to those of the rOspC ELISA and the Fla ELISA for relevant controls including patients with syphilis and mononucleosis. Although the overall diagnostic sensitivity of the Fla ELISA was superior, 8.8 and 12.0% of the EM and NB patients, respectively, were antibody positive only by the pepC10 ELISA. Thus, use of a diagnostic test for LB based on the detection of IgM antibodies to pepC10 and Fla has increased sensitivity for the diagnosis of early LB.

Adolescent↗

PubMed1998

Molecular typing of Borrelia burgdorferi sensu lato by randomly amplified polymorphic DNA fingerprinting analysis.

To study whether pathogenic clusters of Borrelia burgdorferi sensu lato strains occur, we typed 136 isolates, cultured from specimens from patients (n = 49) with various clinical entities and from ticks (n = 83) or dogs (n = 4) from different geographic regions, by randomly amplified polymorphic DNA (RAPD) fingerprinting with four arbitrary primers. The RAPD patterns were reproducible up to the 95% similarity level as shown in duplicate experiments. In these experiments the purified DNAs prepared on different days, from different colonies, and after various passages were used as templates. With an intergroup difference of 55%, the 136 strains could be divided into seven genetic clusters. Six clusters comprised and corresponded to the established species B. burgdorferi sensu stricto (n = 23), Borrelia garinii (n = 39), Borrelia afzelii (n = 59), Borrelia japonica (n = 1), Borrelia valaisiana (n = 12), and genomic group DN127 (n = 1). One strain from a patient with erythema migrans (EM) did not belong to any of the species or genomic groups known up to now. The RAPD types of B. burgdorferi sensu stricto, B. garinii, and B. afzelii isolates, which may give rise to human Lyme borreliosis (LB), were associated with their geographic origins. A high degree of genetic diversity was observed among the 39 B. garinii strains, and six subgroups could be recognized. One of these comprised eight isolates from patients with disseminated LB only and no tick isolates. B. afzelii strains from patients with EM or acrodermatitis chronica atrophicans were not clustered in particular branches. Our study showed that RAPD analysis is a powerful tool for discriminating different Borrelia species as well as Borrelia isolates within species.

Animals↗

PubMed1998

Diagnostic value of PCR for detection of Borrelia burgdorferi in skin biopsy and urine samples from patients with skin borreliosis.

Skin biopsies of 36 patients with erythema migrans and acrodermatitis chronica atrophicans (ACA) before therapy and those of 8 patients after therapy were examined for Borrelia burgdorferi DNA by PCR. Skin biopsies of 27 patients with dermatological diseases other than Lyme borreliosis and those of 10 healthy persons were examined as controls. Two different primer sets targeting 23S rRNA (PCR I) and 66-kDa protein (PCR II) genes were used. PCR was performed with freshly frozen tissue (FFT) and paraffin-embedded tissue (PET). For FFT specimens of erythema migrans, 73% were positive by PCR I, 79% were positive by PCR II, and 88% were positive by combining PCR I and II. For PET specimens, PCR was less sensitive (PCR I, 44%; PCR II, 52%). For FFT specimens of ACA, PCR I was positive for two of five patients and PCR II was positive for four of five patients. B. burgdorferi was cultured from 79% of the erythema migrans specimens but not from any of the ACA lesions. Elevated B. burgdorferi antibodies were detected in sera of 74% of erythema migrans patients and 100% of ACA patients. All urine samples were negative by PCR II, whereas PCR I was positive for 27%. However, hybridization of these amplicons was negative. Sequencing of three amplicons identified nonborrelial DNA. In conclusion, urine PCR is not suitable for the diagnosis of skin borreliosis. A combination of two different primer sets achieves high sensitivity with skin biopsies. In early erythema migrans infection, culture and PCR are more sensitive than serology.

Adult↗

PubMed2001

Chronic borreliosis presenting with morphea- and lichen sclerosus et atrophicus-like cutaneous lesions. a case report.

We report on a case of chronic cutaneous borreliosis with manifestations clinically compatible with morphea and lichen sclerosus et atrophicus. The histopathologic features of these lesions were those of acrodermatitis chronica atrophicans. Our case illustrates the concept that clinical aspects of morphea and lichen sclerosus et atrophicus pertain to the spectrum of cutaneous borreliosis.

Abdomen↗

PubMed2005

Lyme borreliosis: insights into tick-/host-borrelia relations.

Lyme borreliosis (LB) is a serious infectious disease of humans and some domestic animals in temperate regions of the Northern Hemisphere. It is caused by certain spirochetes in the Borrelia burgdorferi sensu lato (s.l.) species complex. The complex consists of 11 species (genospecies). Borrelia burgdorferi sensu stricto (s.s.), Borrelia garinii and Borrelia afzelii are the major agents of human disease. Borrelia burgdorferi s.l. species are transmitted mainly by ticks belonging to the Ixodes ricinus species complex plus a few additional species not currently assigned to the complex. B. burgdorferi infections may produce an acute or chronic disease with a wide array of clinical symptoms such as erythema migrans (EM), carditis, arthritis, neuroborreliosis, and acrodermatitis chronica atrophicans (ACA). Differences in LB spirochetes 'genospecies' and strains/isolates determine the occurrence and severity of this multi-system disease. Accurate and reliable identification of the LB spirochetes in ticks as well as knowledge of their prevalence are essential for prevention against the disease and development of an effective vaccine. An overview of the knowledge of molecular factors with emphasis on potential protein-carbohydrate interactions in the tick-borrelia system is the main focus of this review.

Animals↗

PubMed1990

Prevalence of tick-borne encephalitis and Lyme borreliosis in a defined Swedish population.

Sera from 346 individuals living on Lisö, an area south of Stockholm, endemic for tick-borne encephalitis (TBE) and Lyme borreliosis, were tested for antibody activity to TBE-virus (TBEV) and Borrelia burgdorferi, using a sonicate antigen, by haemagglutination-inhibition and ELISA, respectively. 10/346 (3%) individuals had a history of previous serologically confirmed TBE with encephalitic symptoms, and 33/346 (10%) had a history of previous erythema chronicum migrans (ECM). Four individuals (1%) had been treated for neuroborreliosis and another 4 (1%) for acrodermatitis chronica atrophicans (ACA). Antibodies to TBEV and B. burgdorferi were detected in 40/346 (12%) and 89/346 (26%) individuals, respectively. The seroprevalence of Lyme borreliosis increased with age, time spent on Lisö, and number of reported tick-bites. For TBE there was a correlation between seropositivity and time spent on Lisö only. In sera from 50 healthy blood-donors, living in a non-endemic area, no antibody activity to TBEV could be detected and only 1/50 (2%) had antibodies to borrelia. In sera from 150 age and sex matched control individuals, living in the city of Stockholm, antibody activity to TBEV and borrelia was found in 8/150 (5%) and 13/150 (9%), respectively.

Adolescent↗

PubMed1996

Borrelia burgdorferi sensu lato strains isolated from cutaneous Lyme borreliosis biopsies differentiated by pulsed-field gel electrophoresis.

25 skin biopsy isolates of Borrelia burgdorferi sensu lato, mainly from German patients with erythema migrans [EM], borrelial lymphocytoma [BL] and acrodermatitis chronica atrophicans [ACA], were species-differentiated using pulsed-field gel electrophoresis (PFGE). The isolates revealed 15 B. afzelii, 8 B. garinii and 2 B. burgdorferi sensu stricto species according to Miu I digestion. All 6 ACA isolates were identified as B. afzelii species, whereas the 2 borrelial lymphocytoma isolates were grouped into B. garinii species. 12 B. afzelii strains, including 4 ACA isolates, were further investigated by digestion with 5 restriction enzymes. Most of the strains revealed an individual pattern. No specific characteristically large restriction fragment pattern (LRFP) could be detected within the ACA and/or EM group. The ACA isolates could not be clearly differentiated from the EM isolates according to the LRFP patterns. PFGE is thus a highly effective method for detecting differences among B. afzelii isolates.

Biopsy↗

PubMed1999

Principles of the diagnosis and antibiotic treatment of Lyme borreliosis.

Clinical signs and symptoms are an essential aspect of the diagnosis of Lyme borreliosis. Thus, a thorough knowledge of the clinical features of the disease is important. Established clinical definitions could be of help in everyday clinical practice and especially to compare the findings of different authors or groups. The characteristic sign that permits the diagnosis of Lyme borreliosis is a typical erythema migrans skin lesion. Highly suggestive manifestations are ear lobe lymphocytoma, acrodermatitis chronica atrophicans and Bannwarth's syndrome. The majority of other signs and symptoms are only suggestive and, when expressed individually, may have a very limited or even symbolic value for the purpose of diagnosis. Laboratory confirmation of borrelial infection is needed, as a rule, for all manifestations of Lyme borreliosis with the exception of typical erythema migrans. In clinical practice indirect laboratory methods are usually employed. Determination of borrelial IgM and IgG antibodies by immunofluorescence assays or enzyme-linked immunosorbent assays has not been standardised, and correlation of the results from different laboratories and/or different commercial tests may be poor. Immunoblotting may solve some of the many dilemmas but could (especially in Europe) raise additional questions in a field in which numerous uncertainties already exist. The reliability of methods for direct detection of borrelial infection other than culture to ascertain spirochetes in tissue specimens is open to question. Treatment with antibiotics is reasonable in all stages of Lyme borreliosis and for all clinical manifestations; however, it has been most effective early in the course of the illness. The choice of antibiotic depends upon many factors including the efficacy, pharmacokinetic profile, side effects, expected compliance and price. For the majority of manifestations the most effective antibiotic, the optimal dosage, and the most appropriate duration of treatment have not been exactly determined. Recommendations for the treatment of Lyme borreliosis in Slovenia are presented.

Adult↗

PubMed1999

The outcome of Lyme borreliosis in children.

Austria is an endemic area for Lyme borreliosis. The IgG seroprevalence of healthy blood donors as investigated by a DAKO flagellum-ELISA in Graz/Styria is 13%. In order to determine whether this high seroprevalence is caused by infection in childhood, 36 children aged 3 to 18 years (mean, 10.1 years) were followed up over 2 to 20 months (mean, 11.1 months) and reinvestigated for clinical symptoms and antibodies against B. burgdorferi by a commercial flagellum ELISA and a commercial B. garinii Western blot (WB). Twenty-seven children had erythema migrans (EM), one of them with reinfection, 5 had borrelia lymphocytoma (BL), 2 EM and BL, 1 acrodermatitis chronica atrophicans and 1 ACA/circumscribed scleroderma. Before treatment with either phenoxymethylpenicillin, amoxicillin, or minocyclin for 3-4 weeks, 64% of the patients were IgM and 44%, IgG seropositive. Clinically, all but 5 patients with EM recovered from Lyme borreliosis. Among these 5 patients--one of them with reinfection of EM--3 had mild arthralgia, 1 recurrent headache and 1 concentration disturbance. Only 2 children with arthralgia were IgM positive by ELISA and WB. One of 5 BL patients had a persistent swelling of the ear lobe although with a negative serology before and after several antibiotic treatments and at follow up. In 16 children serological investigations were performed after more than 12 months (range, 13-20 months). Eighteen percent of them had IgM antibodies by ELISA, 25% by WB, and 6% IgG antibodies by ELISA and 6% by WB. Although there was a decline of antibody response from 64% to 18% for IgM and from 44% to 6% for IgG as measured by ELISA, children remain seropositive for more than 1 year with or without clinical symptoms. The relevance of the association with clinical symptoms can be raised by combining several diagnostic methods. It is assumed that recurrent, often silent, infections might increase antibody titres. It should be noted that antibody titres also generally increase with the age of individuals.

Adolescent↗

PubMed2000

[Skin manifestations of Lyme borreliosis in patients at the 1st Dermato-venereologic Clinic of Comenius University Medical School in Bratislava 1996-1998].

60-70% of Lyme borreliosis is formed by early manifestations, in particular erythema migrans (EM). All vegetative forms of the tick Ixodes ricinus (e.g. full grown tick, nymphae and larvae) transmit the causal organism, the spirochete Borrelia burgdorferi sensu stricto and Borrelia burgdorferi sensu lato (B. garinii, B. afzelii). The objective of the work was to evaluate clinical and epidemiological parameters and to contribute by testing and investigating in greater detail criteria to the early and exact diagnosis of the disease. In a group of 50 patients the time of development of EM was evaluated, the mode of transmission, the incubation period, localization, symptoms of dissemination of B. burgdorferi, the period of treatment and the antibiotic used. The dynamics of the antibody titre against B. burgdorferi (IFA test) were assessed in 21 patients with EM for a period of 1 to 22 months and in 5 patients with acrodermatitis chronic atrophicans (ACA) for 3 to 55 months. In 50% EM developed during the summer months. 66% reported as the vector a tick, 14% insects and 20% did not know. An incubation of 1 and 2 weeks was reported in 34%, the lower extremities were affected in 52%. Manifestations of dissemination were found in 6 patients, in 2 patients EM relapsed. Antibodies (Ab) against B. burgdorferi were present in 38 patients. In 21 patients the dynamics of the antibody titre were followed up for 1 to 22 months and no substantial changes were found. Serum positivity in patients with ACA persisted without change of the titre for several years. In the treatment of EM most frequently doxycycline was administered for two weeks. EM as the early stage of LB is a seasonal disease with a natural focus. If treatment is started in the early stage of infection, antibodies against B. burgdorferi need not develop. There is no correlation between clinical complaints and serological results and the type of treatment. The prognosis of the disease is favourable in the majority of patients. Vaccination offers new possibilities in active protection against Borrelia infection, in particular in endemic areas. For European countries at present a recombined vaccine is being prepared from the surface lipoprotein A (OspA) made from prevalent strains of B. afzelii and B. garinii.

Antibodies, Bacterial↗

PubMed2001

[Culture of Borrelia burgdorferi sensu lato from patients in the Czech Republic].

Spirochete of Borrelia burgdorferi sensu lato were isolated in a modified BSK medium from 26 patients of 275 investigated suffering from early and late borreliosis. The isolates were specified by phenotype and genotype analysis using monoclonal antibodies, by immunochemical methods and by including species specific 16S rRNA, OspA and OspC primers and probes in a polymerase chain reaction with hybridization. Borrelia afzelii was found in 9 patients, Borrelia garinii in 16 and B. burgdorferi sensu stricto in one instance. B. afzelii was isolated from 5 biopsies with erythema migrans. 2 with acrodermatitis chronica atrophicans and from blood of 2 patients with arthritis. B. garinii cultures were prepared from 6 cerebrospinal fluids, 3 blood samples, 1 placenta, 1 liver biopsy and 5 skin samples. Unique was B. burgdorferi detected in a bioptic sample of the heart muscle which was positive on immunohistochemical examination. Monoclonal antibodies against OspA and genotype analysis provided evidence that Borrelia garinii isolates from cerebrospinal fluid are close to serotype IV. One CSF isolate was resistant to antibiotics; the tropism of B. garinii to nervous tissue is contemplated.

Adolescent↗

PubMedDate not supplied

[Clinico-epidemiologic characteristics of Lyme disease treated at the Infectious Disease in Novy Sad 1993-1998].

INTRODUCTION: Borrelia burgdorferi, the etiological agent of Lyme disease, is transmitted by the bite of Ixodes ricinus, registered in all parts of Yugoslavia. Vectors are very active in spring and early summer and the disease has a seasonal distribution. Generally speaking, there are three defined stages of the disease, but some of them can be misdiagnosed or really absent. Serological analysis of Lyme disease is very difficult to interpret, especially in later stages, so confirmation by immunoblot assays is recommended. The aim of this study was to present some epidemiologic and clinical characteristics of Lyme disease in Vojvodina in the period from 1993-1998. Throughout this period, 1.659 persons with tick bite were registered, whereas 560 with diagnosed Lyme disease have been treated at the Clinic for Infectious Diseases in Novi Sad. RESULTS: In 511 patients (91.25%) we registered the first stage of the disease, in 42 (7.50%) the second stage and in 7 (1.25%) the third stage of the disease. The mean age of patients with erythema migranes was 38.67 years, mean incubation period was 9.37 days, and tick was removed from the skin after 2.29 days on average. Most of the identified tick bites originated from suburban areas (50.29%), they predominantly occurred in May and June (63.01%), and most of the ticks were removed improperly (57.67%). Dominant clinical manifestations of the second stage were acute meningitis (9.52%), Bannwarth's syndrome (9.52%), arthralgia and arthritis (50%), skin lesions (14.28%), cardiac disorders (11.90%) and mild liver lesions (2.38%) and generalized lymphadenopathy (2.38%). Chronic neuroborreliosis (42.85%), acrodermatitis chronica atrophicans (28.57%) and chronic arthritis were dominant clinical manifestations of the third stage. Up to 81.63% of patients with late stage of disease had a history of previous tick bite. One third of patients were asymptomatic in the first stage of the disease. Improper treatment of the first stage resulted in development of late stage disease in 57.14% of patients. CONCLUSIONS: Morbidity of Lyme disease in Vojvodina is about 1.98-9.8 cases on 100.000 inhabitants, and it belongs to regions with low incidence. Majority of bites are registered during summer months arround cities. Longer persistence of vector on the skin is a risk factor for manifested diseases. Inappropriately treated or completely untreated persons have higher risk for disseminated infection. In our country, most common manifestations of the second stage are neurological and articular manifestations as in the third stage. Leading symptoms and epidemiology aren't enough for diagnosis of Lyme disease, and more specific and sensitive serologic assays are necessary.

Adolescent↗

PubMed2002

Solitary borrelial lymphocytoma in adult patients.

During the period from 1986 to 2000, 85 adult patients with solitary borrelial lymphocytoma were diagnosed at the Department of Infectious Diseases, University Medical Centre Ljubljana, Slovenia. There were 36 (42.4%) females and 49 (57.6%) males with a median age of 49 (15-74) years. Borrelial lymphocytoma was located on the breast (nipple--areola mammae region) in 68 (80%) patients, on the ear lobe in eight (9.4%), and in other locations in nine (10.6%). A concomitant erythema migrans enabling clinical diagnosis of Lyme borreliosis was registered or reported in 67 (78.8%) patients. Fifteen (17.6%) patients had no accompanying symptoms, 34 (40%) reported local and constitutional symptoms, 23 (27.1%) recounted only local symptoms, and 13 (15.3%) patients had solely constitutional symptoms. Clinical findings indicating early disseminated borrelial infection were observed at the first visit in 12 (14.1%) patients: six (7.1%) had multiple erythema migrans, one had meningitis, one meningoradiculitis and arthritis, one radiculoneuritis and arthritis, one peripheral facial palsy and concomitant meningitis, and two arthritis. In addition, one of the patients with borrelial lymphocytoma on the breast had acrodermatitis chronica atrophicans. A seropositive response to borrelial antigens was found in 30 (35.3%) patients at the initial examination. In 11/46 (23.9%) patients, infection with Borrelia burgdorferi sensu lato was confirmed by isolation of the agent from lymphocytoma tissue. Eight out of nine (88.9%) typed borrelial strains were found to be B. afzelii, and one (11.1%) B. bissettii. Patients were treated with doxycycline, azithromycin, amoxycillin, cefuroxime-axetil, phenoxymethylpenicillin, or ceftriaxone. Median time to complete disappearance of lymphocytoma was 28 days (range 7-270 days) after the institution of antibiotic treatment; disappearance took longer in patients with prolonged duration of the skin lesion prior to treatment. Treatment failure was registered in 11 (12.9%) patients who were later re-treated. The outcome of borrelial infection assessed at the end of a follow-up period of one year was favourable.

Adolescent↗

PubMedDate not supplied

Anetoderma and borreliosis: is there a pathogenetic relationship?

A 32-year-old man simultaneously developed anetoderma and acrodermatitis chronica atrophicans on his left arm and showed a positive serology for borreliosis with ELISA and Western Blot tests. In addition, a 45 year-old man is presented with anetoderma without any associated systemic or cutaneous diseases, with B. afzelii confirmed as a singular causality through serology (ELISA, Western Blot) and amplification of B. afzelii-specific DNA from the skin by PCR. These two observations highly suggest that anetoderma can be the result of an infection with B. afzelii. We conclude that in patients with anetoderma a serological investigation for Borreliosis should be performed.

Adult↗

PubMed1952