Diagnosis Of Lyme Borreliosis Research Articles

Production, purification, and quality assessment of borrelial proteins CspZ from Borrelia burgdorferi and FhbA from Borrelia hermsii

Borrelia, spirochetes transmitted by ticks, are the etiological agents of numerous multisystemic diseases, such as Lyme borreliosis (LB) and tick-borne relapsing fever (TBRF). This study focuses on two surface proteins from two Borrelia subspecies involved in these diseases: CspZ, expressed by Borrelia burgdorferi sensu stricto (also named BbCRASP-2 for complement regulator-acquiring surface protein 2), and the factor H binding A (FhbA), expressed by Borrelia hermsii. Numerous subspecies of Borrelia, including these latter, are able to evade the immune defenses of a variety of potential vertebrate hosts in a number of ways. In this context, previous data suggested that both surface proteins play a role in the immune evasion of both Borrelia subspecies by interacting with key regulators of the alternative pathway of the human complement system, factor H (FH) and FH-like protein 1 (FHL-1). The recombinant proteins, CspZ and FhbA, were expressed in Escherichia coli and purified by one-step metal-affinity chromatography, with yields of 15 and 20 mg or pure protein for 1 L of cultured bacteria, respectively. The purity was evaluated by SDS-PAGE and HPLC and is close to about 95%. The mass of CspZ and FhbA was checked by mass spectrometry (MS). Proper folding of CspZ and FhbA was confirmed by circular dichroism (CD), and their biological activity, namely their interaction with purified FH from human serum (recombinant FH15-20 and recombinant FHL-1), was characterized by SPR. Such a study provides the basis for the biochemical characterization of the studied proteins and their biomolecular interactions which is a necessary prerequisite for the development of new approaches to improve the current diagnosis of LB and TBRF.Key points• DLS, CD, SEC-MALS, NMR, HPLC, and MS are tools for protein quality assessment• Borrelia spp. possesses immune evasion mechanisms, including human host complement• CspZ and FhbA interact with high affinity (pM to nM) to human FH and rFHL-1Graphical

The role of clinical characteristics of Lyme borreliosis in predicting cognitive impairment

Background. Lyme disease is characterized by a wide polymorphism of clinical manifestations, including various lesions of the central and peripheral nervous systems. Along with neurological disorders, diverse manifestations and signs of cognitive impairments are often found. Despite the high prevalence of cognitive disorders in Lyme borreliosis, data on their frequency and risk factors are contradictory. The examination of cognitive status is still not included in the standard clinical examination of this category of patients. The aim of the study is to identify predictors that may independently influence the development of cognitive dysfunction in patients with Lyme borreliosis. Materials and methods. A prospective cohort study of 69 patients (47 females, 22 males) diagnosed with Lyme borreliosis, aged between 23 and 77 (average of 49.90 ± 16.26) years, was carried out. The Montreal Cognitive Assessment was used to evaluate cognitive function. To determine the relationship between cognitive dysfunction and its potential predictors, the participants were divided into two groups: patients without cognitive impairment (n = 30) and those with cognitive dysfunction (n = 39) based on the Montreal Cognitive Assessment score. Results. It was found that risk factors for the development of cognitive impairment include: diagnosis of Lyme borreliosis at the age of 45.5 years and older (hazard ratio (HR) 5.09; 95% confidence interval (CI) [1.82–14.27]; p = 0.001), presence of neuroborreliosis (HR 5.98; 95% CI [2.0–17.8]; р < 0.001), Lyme carditis (HR 8.7; 95% CI [1.04–73.06]; р = 0.021), hypertension (HR 7.19; 95% CI [2.32–22.28]; р < 0.001), anxiety disorders. Conclusions. The prospect for further research is to study and analyze the features of the correlation between cognitive and psycho-emotional disorders and the quality of life of patients with different forms and duration of Lyme borreliosis.

Discrepancy between IDSA and ESGBOR in Lyme disease: Individual participant meta-analysis in Türkiye.

The evidence on the prevalence of Lyme borreliosis (LB) is limited, but there is a suspicion of overdiagnosis of LB in recent years. We reviewed the LB diagnosis and treatment-related data in Türkiye, based on the Infectious Diseases Society of America (IDSA) 2020 and European Society of Clinical Microbiology and Infectious Diseases Study Group for Lyme Borreliosis (ESGBOR) 2018 guidelines. By detecting the disagreements between these two, we outlined the areas to be improved for future guidelines. We performed a literature search according to the PRISMA guidelines in PubMed, Ovid-Medline, Web of Science, Turkish Medline, Scopus, CINAHL, ULAKBIM TR Index, Google Scholar and Cochrane Library databases. We included the published cases in a database and evaluated according to IDSA and ESGBOR guidelines. We outlined the reasons for misdiagnoses and inappropriate uses of antibiotics. We included 42 relevant studies with 84 LB cases reported from Türkiye between 1990 and December 2022. Among 84 cases, the most common clinical findings were nervous system findings (n = 37, 44.0%), erythema migrans (n = 29, 34.5%) and ophthalmologic findings (n = 15, 17.9%). The IDSA 2020 and ESGBOR 2018 guidelines agreed on the diagnosis of 71 (84.5%) cases; there was an agreement that 31 cases (36.9%) were misdiagnosed and 40 cases (47.6%) were correctly diagnosed, and there was disagreement for 13 cases (15.5%). Serum immunoglobulin M (IgM), IgG measurements by ELISA and western blot were widely performed, and they were effective in definitive diagnosis merely when used according to guidelines. Inappropriate use of antibiotics was detected in 42 (50.0%) of cases which were classified in the following categories: incorrect LB diagnosis, inappropriate choice of antibiotic, inappropriate route of drug administration and prolonged antibiotic treatment. Overdiagnosis and non-adherence to guidelines is a common problem. The discordance between seroprevalence and clinical studies necessitates a consensus over the best clinical approach.

Adaptive Immune Response Investigation in Lyme Borreliosis.

To diagnose Lyme Borreliosis, it is advised to use an enzyme-linked immunosorbent test to check for serum antibodies specific for Lyme and all tests with positive or ambiguous enzyme-linked immunosorbent assay (ELISA) results being confirmed by immunoblot. This method of measuring the humoral immunity in human fluids (e.g., by ELISA) has provided robust and reproducible results for decades and similar assays have been validated for monitoring of B cell immunity. These immunological tests that detect antibodies to Borrelia burgdorferi are useful in the diagnosis of Borreliosis on a routine basis. The variety of different Borrelia species and their different geographic distributions are the main reasons why standards and recommendations are not identical across all geographic regions of the world. In contrast to humoral immunity, the T cell reaction or cellular immunity to the Borrelia infection has not been well elucidated, but over time with more studies a novel T cell-based assay (EliSpot) has been developed and validated for the sensitive detection of antigen-specific T cell responses to B. burgdorferi. The EliSpot Lyme assay can be used to study the T cell response elicited by Borrelia infections, which bridges the gap between the ability to detect humoral immunity and cellular immunity in Lyme disease. In addition, detecting cellular immunity may be a helpful laboratory diagnostic test for Lyme disease, especially for seronegative Lyme patients. Since serodiagnostic methods of the Borrelia infection frequently provide false positive and negativeresults, this T cell-based diagnostic test (cellular assay) may help in confirming a Lyme diagnosis. Many clinical laboratories are convinced that the cellular assay is superior to the Western Blot assay in terms of sensitivity for detecting the underlying Borrelia infection. Research also suggests that there is a dissociation between the magnitude of the humoral and the T cell-mediated cellular immune responses in the Borrelia infection. Lastly, the data implies that the EliSpot Lyme assay may be helpful to identify Borrelia infected individuals when the serology-based diagnostic fails to do so. Here in this chapter the pairing of humoral and cellular immunity is employed to evaluate the adaptive response in patients.

Algorithm for monitoring patients after tick bite in the context of Lyme-borreliosis

Timely diagnosis of Lyme-borreliosis (LB) is the key to its successful treatment. Outpatient monitoring of patients after tick bite based on the algorithm is able to optimize the early diagnosis of LB, shorten the start of treatment and improve the disease surveillance.Purpose. Creation of an algorithm for monitoring patients after tick bite for use in outpatient settings by primary care physicians. Material and methods. A retrospective, non-randomized, single-center cohort study was conducted based on the analysis of data from 660 outpatient records of patients consulted by an infectious disease specialist in the Infectious clinical hospital No.1 in Moscow. Patients were divided by age and the presence or absence of a diagnosis of LB. Confirmation of the diagnosis was carried out according to clinical and/or laboratory criteria using methods of enzyme-linked immunoassay and immune blot. Results. The analysis of the results of enzyme immunoassay of antibodies to borrelia in patients with LB was carried out. The optimal sampling time was 7 or more weeks from the moment of tick bite and 4 or more weeks from the moment of onset of symptoms. The specificity of immunological diagnostics in patients with excluded LB was evaluated. The proportion of doubtful and false positive results in such patients was high — 28.1%. False positive results were more common in adults (37.6%) compared with children (9.8%), which is associated with more frequent false positive results of IgM determination (34.1% in adults, 4.9% in children). Based on the data of our study, an algorithm for monitoring patients after tick bite has been developed, focused on use in outpatient settings by primary care physicians.

Is there Lyme borreliosis in French Guiana? Descriptive study among patients referred for a suspected Lyme borreliosis in an Amazonian hospital between 2010 and 2022

Lyme borreliosis (LB) existence in South America is debated, especially in the Amazon region. The infection with Lyme borreliae has never been reported in French Guiana where Borrelia burgdorferi sensu lato is not found in ticks. We describe the final diagnosis and presumed place of acquisition in patients consulting for suspicion of LB.We retrospectively collected data from all consecutive patients consulting for a suspicion of LB between 2010 and 2021 at Cayenne Hospital, French Guiana. Patients were classified by an adjudication committee as confirmed LB if they met the criteria of the French consensus, as possible LB if they had compatible symptoms and a good outcome after appropriate treatment, or excluded when a differential diagnosis was found. The place of acquisition was discussed in case of possible or confirmed case.Twenty-six patients were included. Rheumatologic symptoms were the most reported (88 %) followed by neurological symptoms (61 %). Twenty-four (92 %) of these patients were born out of French Guiana. Diagnosis of LB was considered as confirmed in 2 patients (8 %), for whom the place of acquisition was likely mainland France, and as possible in 3 patients (11 %) with early localized LB presumably acquired in French Guiana. Functional somatic disorders were diagnosed in 13 (50 %) patients whereas 9 (35 %) were found with another disease.This study did not confirm the acquisition of LB in French Guiana. However, three possible autochthonous cases encourage clinicians working in the Amazon area to stay aware of LB.

Knowledge, protection behaviours and seroprevalence of Lyme borreliosis in inhabitants of Lublin Province, eastern Poland - evaluation of a prophylaxis programme.

Lyme borreliosis (LB) is the most frequent tick-borne disease with 17,338 cases reported in Poland in 2022. Since research on a LB vaccine is still ongoing, the promotion of individual behaviours and limiting of tick exposure is one of the most effective ways to prevent LB. The aim of the study was to evaluate the effectiveness of the LB prevention programme by assessing the knowledge, practice behaviours, seroprevalence of LB and satisfaction among the population of the Lublin Province in eastern Poland. The prevention programme was carried out among 2,920 participants who were asked about their exposure to ticks, history of LB and prevention behaviours. Awareness of knowledge was evaluated before and after training. Seroprevalence of LB was rated by ELISA and immunoblot assays. Over 73% of participants reported tick bites in their lifetime, without significant differences between rural and urban area inhabitants. More than 80% of individuals declared that they use protective measures (PPM), such as proper clothes and body checking; repellents were the least frequently used by participants. The diagnosis of LB but not tick bites in a lifetime influenced the more frequent use of PPM. Increase in knowledge was observed in 86% of participants after education, and the highest knowledge was noted among respondents with higher education. The seroprevalence of anti-B.burgdorferi antibodies was 37% and was higher among men than women (40% vs. 36%). The population of Lublin Province is highly exposed to tick bites and infection with B. burgdorferi. The high seroprevalence and increase in knowledge confirmed the effectiveness and need for preventive programmes. These results can be useful for optimizing and enhancing the effects of future prevention campaigns.

Citizen Science on Lyme Borreliosis in Spain Reveals Disease-Associated Risk Factors and Control Interventions.

Background: Lyme borreliosis (LB) caused by Borrelia burgdorferi sensu lato complex spirochetes is one of the tick-borne diseases with high prevalence and social/economic burden in the United States, Spain, and other European countries. The objective is to address limited information available about the incidence, prevalence, and symptoms of LB, current prevention, and treatment interventions that are not adequately focused and thus not very effective against this disease. Methods: To address these limitations, in this study, we used a citizen science approach to evaluate the LB-associated risks and implementation of control interventions in Spain. A total of 405 participants in the survey were included in the analysis. Responses to the questionnaire were received during January-July 2022. The questionnaire contained qualitative and quantitative questions. Homogeneity among binary variables was analyzed using a Fisher's exact test. Results: Despite limitations of the study associated with response to the questionnaire and information on tick species, the results evidenced the effect of factors such as age, gender, tick bites, disease clinical signs, comorbidities such as alpha-gal syndrome, health care services, and treatment effectiveness affecting LB. Conclusions: The main conclusions of the study highlight the need for better surveillance of tick infestations, pathogen infection, and diagnosis of LB and related comorbidities. To advance in disease prevention, diagnosis, and treatment, new interventions need to be developed and implemented in both public and private health care services. Providing access to these results to the society, health care system, and scientists is important to further advance in disease surveillance, diagnosis, control, and prevention.

Utility of Borrelia-specific IgM and IgG antibody titer determinations during a 12-year period - results from a clinical laboratory in Northern Sweden.

Interpretation of serological findings in suspected Lyme borreliosis (LB) is challenging and IgM reactivities may have low predictive value. Therefore, if used indiscriminately, there is a risk for incorrect diagnosis of LB. To evaluate the usefulness of IgM titer determination, we performed a study of the prevalence of Borrelia-specific antibodies in serological samples from patients with suspected LB analyzed during the period 2010 - 2021 at the University Hospital of Umeå in Sweden. In total, 19,335 samples had been analyzed for the presence of IgG and IgM antibodies. Overall, there were higher percentages of IgM positive or borderline titers, 1,847 (9.6%) and 905 (4.7%), respectively, than IgG positive or borderline titers, 959 (5.0%) and 406 (2.1%), respectively. Peak number of samples were recorded 2012 - 2013, exceeding 1,800, whereas there were around 1,200 during 2020 - 2021. The peak number of positive IgG and/or positive IgM samples were observed during the period 2015 - 2017 with close to, or above 400, and concomitantly, the proportion of IgG positive samples increased markedly. For IgG positive samples, the increase followed a positive linear time trend (P< 0.001). Peak monthly numbers were observed during August, September, and October. This seasonal increase was significant for the IgG positive group (P< 0.05), but not for the IgM positive/IgG negative group. Repeated samples were obtained from 3,188 individuals and of the initial samples 2,817 were (88%) IgG negative and 2,315 (72%) were IgM negative and of these, 130 (4%) showed IgG seroconversion and 300 (9%) IgM seroconversion. Collectively, the data demonstrate that IgG and/or IgM positive samples represented a minority of all samples, even when repeated sampling had occurred, and IgM positive samples were much more common than IgG positive samples. Thus, the accuracy of the clinical suspicion was low and this will lead to a low predictive value of the analysis, in particular of IgM. These findings question the use of IgM titer determination as a routine analysis.

The history of Lyme disease in Italy and its spread in the Italian territory.

Lyme borreliosis (LB) is the most common vector-borne zoonotic inflammatory disease in the Northern Hemisphere. In Italy, the first case was diagnosed in 1985 in a woman in Liguria, while the second, in 1986 in Friuli-Venezia Giulia, documenting the infection in northern Italy. Both diagnoses were confirmed by serological assessment by an indirect immunofluorescence (IFI) technique. Borrelia cultivation from both Ixodes ricinus ticks and human lesions in Trieste (Friuli-Venezia Giulia) identified Borrelia afzelii as the prevalent genospecies; nevertheless, Borrelia garinii, Borrelia burgdorferi (sensu stricto), and Borrelia valaisiana (VS116 Group) were also detected, although less frequently. LB was also documented in other Italian regions: in Tuscany (1991), Trentino-Alto Adige (1995-1996), Emilia-Romagna (1998), Abruzzo (1998), and more recently, Lombardy. Nevertheless, data on LB in other Italian regions, especially in southern Italy and islands, are poor. The aim of this study is to document the spread of LB in Italy through the collection of data from LB patients in eight Italian hospitals located in different Italian regions. Diagnostic criteria for LB diagnosis are as follows: i) the presence of erythema migrans (EM) or ii) a clinical picture suggestive of LB, confirmed by serological tests and/or PCR positivity for Borrelia detection. In addition, data also included the place of residence (town and region) and the place where patients became infected. During the observation period, 1,260 cases were gathered from the participating centers. Although different in extent from northern Italy to central/southern Italy, this study shows that LB is widespread throughout Italy.

Seroprevalence of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum Infections in German Horses.

There are limited data on Lyme borreliosis (LB), a tick-borne disease caused by the Borrelia burgdorferi sensu lato complex, in horses. Seropositivity is not necessarily associated with clinical disease. Data on seropositivity against Borrelia burgdorferi and Anaplasma phagocytophilum in German horses are sparse. Therefore, serum samples from horses (n = 123) suspected of having Lyme borreliosis and clinically healthy horses (n = 113) from the same stables were tested for specific antibodies against Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. The samples were screened for antibodies against Borrelia burgdorferi (ELISA and an IgG line immunoblot assay). Furthermore, the samples were examined for antibodies against B. burgdorferi and Anaplasma phagocytophilum with a validated rapid in-house test (SNAP® 4Dx Plus® ELISA). The clinical signs of suspect horses included lameness (n = 36), poor performance (n = 19), and apathy (n = 12). Twenty-three percent (n = 26) of suspect horses and 17% (n = 18) of clinically healthy horses were seropositive for having a Borrelia burgdorferi sensu lato infection (p = 0.371), showing that the detection of specific antibodies against B. burgdorferi alone is not sufficient for a diagnosis of equine LB. Anaplasma phagocytophilum seropositivity and seropositivity against both pathogens was 20%/6% in suspect horses and 16%/2% in the clinically healthy population, showing only minor differences (p = 0.108). Unspecific testing for antibodies against B. burgdorferi without clinical suspicion of Lyme borreliosis is not recommended since the clinical relevance of seropositivity against Borrelia burgdorferi sensu lato remains to be elucidated.

Diagnostic value of PCR analysis of synovial fluid for the diagnosis of Lyme borreliosis in dogs

Lyme borreliosis, also known as Lyme disease, is a chronic multiorgan disease of humans and animals transmitted by ticks of the Ixodidae family and caused by a group of spirochetes Borrelia burgdorferi sensu lato. Dogs are one of the most susceptible animal species to Lyme disease. Symptoms of the disease in dogs can range from mild lameness and fever to more severe and potentially life-threatening conditions, such as kidney damage and neurological disorders. Due to the variety of clinical manifestations and the lack of a universal diagnostic approach, veterinarians often face difficulties in accurately diagnosing the disease. Therefore, improving the algorithms for diagnosing Lyme borreliosis in dogs remains an urgent issue for practical veterinary medicine. The article describes and analyzes data on two cases of dogs from Kyiv (Ukraine) that presented with musculoskeletal disorders, including lameness, swelling of the knee joints, and tenderness, which raised suspicion of Lyme borreliosis. The diagnostic tests used to confirm the diagnosis of Lyme borreliosis included clinical blood test, C-reactive protein test, cerebrospinal fluid cytology, enzyme-linked immunosorbent assay for IgM and IgG antigens to Borrelia burgdorferi sensu lato, computed tomography and PCR analysis of synovial fluid for the presence of Borrelia burgdorferi sensu lato DNA. As a result, the clinical blood test, C-reactive protein test, and cerebrospinal fluid cytology were not diagnostically important for the diagnosis of Lyme borreliosis. In both cases, serological tests for antibodies to Borrelia burgdorferi sensu lato were not positive. Computed tomography showed the development of an inflammatory process in the knee joints with the formation of osteophytes, enthesophytes, sclerosis of the articular surfaces, and an increase in the volume of synovial bursa. The final diagnosis of Lyme arthritis in both dogs was established by PCR analysis of synovial fluid from the affected joints for the presence of Borrelia burgdorferi sensu lato DNA. This study emphasizes the importance of considering Lyme borreliosis as a potential cause of musculoskeletal disorders in dogs. We also propose an expanded algorithm for the diagnosis of Lyme borreliosis in dogs with a pronounced symptom complex of Lyme arthritis using the method of PCR analysis of synovial fluid to detect the DNA of Borrelia burgdorferi sensu lato spirochetes. Key words: Lyme borreliosis in dogs, clinical cases, diagnosis, polymerase chain reaction, Borrelia burgdorferi sensu lato.

Classical Borrelia Serology Does Not Aid in the Diagnosis of Persistent Symptoms Attributed to Lyme Borreliosis: A Retrospective Cohort Study.

The diagnosis of Lyme borreliosis is based on two-tier testing using an ELISA and Western blot. About 5-10% of patients report persistent symptoms of unknown etiology after treatment, resulting in substantial difficulties in further diagnostic workup. This paper presents a study aimed at determining whether serology can differentiate between patients with persistent symptoms attributed to Lyme and other patients with Lyme borreliosis. A retrospective cohort study included 162 samples from four subgroups: patients with persistent symptoms of Lyme (PSL), early Lyme borreliosis with erythema migrans (EM), patients tested in a general practitioner setting (GP), and healthy controls (HC). ELISA, Western blots, and multiplex assays from different manufacturers were used to determine inter-test variations in PSL and to compare reactivity against Borrelia-specific antigens among the groups. In comparing the IgG and IgM reactivity by Western blot, IgG was more often positive in the PSL group than in the GP group. The individual antigen reactivity was similar between the PSL and EM or GP groups. Inter-test agreement among the manufacturers was variable, and agreement was higher for IgG testing compared to IgM. Serological testing is unable to define the subgroup of patients with persistent symptoms attributed to Lyme borreliosis. Additionally, the current two-tier testing protocol shows a large variance among different manufacturers in these patients.

Incidence of Lyme Borreliosis in the Dutch General Practice Population: A Large-Scale Population-Based Cohort Study Across the Netherlands Between 2015 and 2019

Background: There is a need for updated incidence rates (IRs) of Lyme borreliosis (LB) in Europe, including the Netherlands. We estimated LB IRs stratified by geographic area, year, age, sex, immunocompromised status, and socioeconomic status (SES). Methods: All subjects registered in the PHARMO General Practitioner (GP) Database without prior diagnosis of LB or disseminated LB and having ≥1 year of continuous database enrolment were included. IRs and corresponding confidence intervals (CIs) of GP-recorded LB, erythema migrans (EM), and disseminated LB were estimated during the period 2015‒2019. Results: We identified 14,794 events (suspected, probable, or confirmed) with a diagnostic code for LB that included 8219 with a recorded clinical manifestation: 7985 (97%) with EM and 234 (3%) with disseminated LB. National annual LB IRs were relatively consistent, ranging from 111 (95% CI 106‒115) in 2019 to 131 (95% CI 126‒136) in 2018 per 100,000 person-years. Incidence of LB showed a bimodal age distribution, with peak IRs observed among subjects aged 5‒14 and 60‒69 years in men and women. Higher LB incidence was found in subjects who were residents of the provinces of Drenthe and Overijssel, immunocompromised, or of lower SES. Similar patterns were observed for EM and disseminated LB. Conclusions: Our findings confirm that LB incidence remains substantial throughout the Netherlands with no indication of decline in the past 5 years. Foci in two provinces and among vulnerable populations suggest potential initial target groups for preventive strategies such as vaccination.

Cost of illness in patients with post-treatment Lyme disease syndrome in Belgium.

A proportion of patients with Lyme borreliosis (LB) report long-term persisting signs and symptoms, even after recommended antibiotic treatment, which is termed post-treatment Lyme disease syndrome (PTLDS). Consensus on guidance regarding diagnosis and treatment is currently lacking. Consequently, patients suffer and are left searching for answers, negatively impacting their quality of life and healthcare expenditure. Yet, health economic data on PTLDS remain scarce. The aim of this article is therefore to assess the cost-of-illness related to PTLDS, including the patient perspective. PTLDS patients (N = 187) with confirmed diagnosis of LB were recruited by a patient organization. Patients completed a self-reported questionnaire on LB-related healthcare utilization, absence from work and unemployment. Unit costs (reference year 2018) were obtained from national databases and published literature. Mean costs and uncertainty intervals were calculated via bootstrapping. Data were extrapolated to the Belgian population. Generalized linear models were used to determine associated covariates with total direct costs and out-of-pocket expenditures. Mean annual direct costs amounted to €4618 (95% CI €4070-5152), of which 49.5% were out-of-pocket expenditures. Mean annual indirect costs amounted to €36081 (€31312-40923). Direct and indirect costs at the population level were estimated at €19.4 and 151.5 million, respectively. A sickness or disability benefit as source of income was associated with higher direct and out-of-pocket costs. The economic burden associated with PTLDS on patients and society is substantial, with patients consuming large amounts of non-reimbursed healthcare resources. Guidance on adequate diagnosis and treatment of PTLDS is needed.

1354. Incidence, time trends and geographic distribution of Lyme neuroborreliosis in Denmark using public surveillance data, 2015-2019

Abstract Background Lyme neuroborreliosis (LNB) is a potentially severe manifestation of Lyme borreliosis (LB), a spirochetal tick-borne infectious disease caused by Borrelia burgdorferi. We describe the incidence, time trends and geographic distribution of LNB in Denmark, with the aim to inform public health policy, including the potential use of new vaccines under development. Methods Based on the case definition for LNB (clinical diagnosis of LB plus detection of antibodies in blood and cerebral spinal fluid [confirmed case], OR clinical symptoms compatible with LNB and detection of antibodies in blood [probable case]), we obtained cases reported by laboratories and physicians (2015–2019) from the online platform maintained by the Statens Serum Institut (SSI). The LNB incidence (per 100,000 persons) was calculated by dividing LNB cases by the population data (denominator) obtained from Statistics Denmark. Results Between 2015 and 2019, laboratories reported annually 162 to 200 LNB cases, while physicians reported 48 to 68 cases (Figure). The annual LNB incidence notified by laboratories ranged from 2.8 (95% CI: 2.4‒3.3) to 3.4 (95% CI: 3.0‒4.0) per 100,000 persons for the 5 study years. The average annual LNB incidence per 100,000 persons for 5 Danish regions ranged from 2.3 to 3.3; for 11 provinces, from 1.9 to 7.6; and for 98 municipalities, from 0 to 22.1. Incidence peaks occurred in persons 5‒14 and 65‒74 years of age. Higher incidences were observed among males versus females in all age groups. LNB cases were reported throughout the year, with peaks in July to September. Number of LNB cases and annual incidence (per 100,000 population) notified by (A)physicians and (B) laboratories Conclusion LNB incidence in Denmark is moderate with no evidence of decline. Cases occurred across all regions but were focally concentrated among residents of some municipalities. By analyzing and reporting spatially granular data, we could identify areas with the highest LNB incidence. This may assist policy makers with assessing the efficiency of geographically targeted versus national prevention interventions, including vaccines should these be licensed. To this end, expanding the current surveillance system to include other manifestations of LB would be valuable to better understand geographic endemicity. Disclosures Jozica Skufca, Epidemiologist, p95: Paid by Pfizer to perform the study Nick DeSmedt, Masters in Computer Engineering, P95: P95 was paid by Pfizer to perform the study Andreas Pilz, PhD, Pfizer: Employee|Pfizer: Stocks/Bonds|Pfizer: Stocks/Bonds Andrew Vyse, Ph.D., Pfizer: Stocks/Bonds Elizabeth Begier, M.D., M.P.H., Pfizer: Employee|Pfizer: Stocks/Bonds Maxim Blum, Ph.D., P95: Paid by Pfizer to perform the study Margarita Riera, MD, MPH, P95: Paid by Pfizer to perform the study Bradford Gessner, MD, MPH, Pfizer: Stocks/Bonds Mette Skovdal, Ph.D., Pfizer Denmark: Stocks/Bonds James Stark, Ph.D., Pfizer: Stocks/Bonds.

Detection of antibodies to &lt;i&gt;B. burgdorferi&lt;/i&gt; by enzyme immunoassay in patients with Lyme borreliosis

The detection of antibodies to borrelia by enzyme immunoassay (ELISA) significantly depends on the time from the onset of the disease.Purpose: analysis of the results of antibodies determination to borrelia by ELISA in children and adults with Lyme borreliosis (LB) at various periods from the onset of the disease.Material and methods. We conducted a retrospective, non-randomized, single-center cohort study, based on the analysis of data from 178 outpatient records of patients with a confirmed diagnosis of LB and the presence of antibody detection results by ELISA. Immunological confirmation of the diagnosis of LB was carried out by using ELISA and western blot test systems registered in the territory of the Russian Federation for the separate determination of immunoglobulins to Borrelia burgdorferi antigens of classes M and G. Results. When counting from the date of the onset of the disease, IgM and/or IgG were determined in 76% of patients at 4-6 weeks, and starting from the 7th week – in 95%. When counting from the date of tick bite, IgG with or without IgM was determined in 83% of patients starting from 7th week. At the same time, a significantly large proportion of seronegative patients among children was revealed. We have clarified the duration of antibody persistence after antibacterial therapy. In the interval from 1 to 6 months, antibodies are detected in 73% of patients. For a period of 6 months or more, antibodies can be detected in 42% of patients.Conclusion. The optimal time for detecting antibodies from the disease onset is 4-6 weeks. Antibodies after antibiotic therapy can persist for a long time, in a third of patients up to 6 months or more.

A Novel High Discriminatory Protocol for the Detection of Borrelia afzelii, Borrelia burgdorferi Sensu Stricto and Borrelia garinii in Ticks.

Bacteria of the Borrelia burgdorferi sensu lato complex are the causative agents of Lyme borreliosis (LB). Even if the conventional diagnosis of LB does not rely on the species itself, an accurate species identification within the complex will provide a deepened epidemiological scenario, a better diagnosis leading to a more targeted therapeutic approach, as well as promote the general public's awareness. A comparative genomics approach based on the 210 Borrelia spp. genomes available in 2019 were used to set up three species-specific PCR protocols, able to detect and provide species typing of Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.) and Borrelia garinii, the three most common and important human pathogenic Lyme Borrelia species in Europe. The species-specificity of these protocols was confirmed on previously identified B. afzelii, B. burgdorferi s.s. and B. garinii specimens detected in Ixodes ricinus samples. In addition, the protocols were validated on 120 DNA samples from ticks collected in Sweden, showing 88% accuracy, 100% precision, 72% sensitivity and 100% specificity. The proposed approach represents an innovative tool in epidemiological studies focused on B. burgdorferi s.l. occurrence in ticks, and future studies could suggest its helpfulness in routine diagnostic tests for health care.

СРАВНЕНИЕ КЛИНИЧЕСКОЙ КАРТИНЫ ИКСОДОВОГО КЛЕЩЕВОГО БОРРЕЛИОЗА У ДЕТЕЙ И ВЗРОСЛЫХ В АМБУЛАТОРНЫХ УСЛОВИЯХ

The polymorphism of clinical manifestations of Lyme borreliosis (LB) creates significant problems in the study of this disease. In addition to the variability in classifications used in Russia and abroad, the spectrum and the frequency of symptoms in outpatient and hospitalized patients with LB differ significantly, which makes it difficult for a physician to understand the clinical picture of LB. The purpose of the study was to describe the clinical picture of LB in an outpatient infectologist appointment conditions according to the E. Asbrink, A. Hovmark classification of Lyme borreliosis in children compared with adults. Materials and methods of the research: a retrospective single-center cohort study was conducted based on the analysis of the data from 213 outpatient records of patients with confirmed LB diagnosis, consulted by infectologist at the Moscow Infectious Clinical Hospital No. 1 of the Moscow Department of Healthcare (Moscow, Russia) in Jan. 2018 - Sep. 2019. In order to compare the clinical features between children and adults the patients were divided into 2 groups by age: adults (18 years and older) and children (1 to 17 years old). The confirmation of the LB diagnosis was carried out according to the clinical and/or laboratory criteria using enzyme immunoassay and immune blotting. Results: in the group of children (n=52), the median age was 6 [3; 8] years old, the proportion of boys was 44% (n=23, 95% CI 30-57%), girls - 56% (n=29, 95% CI 43-70%). In the group of adults (n=161), the median age was 59 [49; 65] years old, the portion of males was 29.2% (n=47, 95% CI 22.2-36.0%), females - 70.8% (n=114, 95% CI 64.0-77.8 %). The most frequently diagnosed was LB at its early localized stage, both in children (n=44, 84.0%, 95% CI 74.0-93.0%) and adults (n=123, 76.4%, 95 % CI 69.7-82.6%). No statistically significant difference was found (p=0.248). Erythema migrans (EM) was the vast majority of early localized stage manifestations in children (95%, 95% CI 88-100%). The incidence of EM in adults was not statistically significantly different from that in children (94.3%, 95% CI 89.9-98.1%, p=1.000). EM in children was most often localized on the scalp (46%, 95% CI 31-60%), in adults the most common localizations were the skin of the trunk and lower extremities (total 92.9%, 95% CI 87.8-97.0; n=118). Differences in the typical localization of EM in children and adults were statistically significant (p&lt;0.001). In the clinical picture of the early localized stage of LB the fever was rare in both children (7%, 95% CI 4-16%) and adults (8.9%, 95% CI 4.3-14.5%) (p=1.000). At the same time, the regional lymphadenitis was detected statistically significantly more often in adults than in children (50%, 95% CI 35-65% vs. 26.8%, 95% CI 19.3-34.5%, p=0.005). Both groups showed a lower incidence of regional lymphadenitis during examination by an infectologist if the patient was prescribed antibiotic therapy at the place of residence, while statistically significantly more often in the group of children (34%, 95% CI 23-46 vs. 15%, 95% CI 8-23, p=0.003). Conclusion: the authors have described the spectrum and frequency of the LB clinical manifestations according to the E. Asbrink, A. Hovmark classification in children vs. adults. The influence of research conditions and the previous antibiotic therapy on the spectrum and frequency of the LB clinical manifestations detected during the initial examination was demonstrated as well.

Emergence of Lyme Disease on the French Riviera, a Retrospective Survey

BackgroundThe French Riviera has been declared free of Lyme Borreliosis (LB) for years. Many patients are referred for presumed LB, sometimes with atypical clinical signs and/or doubtful serology, calling the diagnosis into question.MethodsPatients were assessed for LB diagnosis, depending on clinical presentation, laboratory findings, and further examination by other medical professionals.ResultsAmong 255 patients, 45 (18%) were classified as confirmed LB cases [including 28 ongoing LB (10%) and 17 past LB (8%)], and for 210 (82%) a Lyme borreliosis diagnosis was ruled out. Among ongoing LB, 56% had been exposed to or bitten by ticks, exclusively in rural locations of the Alpes-Maritimes. As a result of the diagnostic procedure, 132 (52%) patients had been treated. An alternative diagnosis was established for 134 (52%) patients, covering a wide range of conditions, including mainly psychological (28%) and neurological conditions (25%) or inflammatory and systemic diseases (22%).ConclusionsOur results strongly suggest the endemicity of LB in the Alpes-Maritimes region. Confirmed LB accounted for 18% of patients while 52% were diagnosed with other conditions.