Serologic Tests For Lyme Disease Research Articles

Rapid single-tier serodiagnosis of Lyme disease

Point-of-care serological and direct antigen testing offers actionable insights for diagnosing challenging illnesses, empowering distributed health systems. Here, we report a POC-compatible serologic test for Lyme disease (LD), leveraging synthetic peptides specific to LD antibodies and a paper-based platform for rapid, and cost-effective diagnosis. Antigenic epitopes conserved across Borrelia burgdorferi genospecies, targeted by IgG and IgM antibodies, are selected to develop a multiplexed panel for detection of LD antibodies from patient sera. Multiple peptide epitopes, when combined synergistically with a machine learning-based diagnostic model achieve high sensitivity without sacrificing specificity. Blinded validation with 15 LD-positive and 15 negative samples shows 95.5% sensitivity and 100% specificity. Blind testing with the CDC’s LD repository samples confirms the test accuracy, matching lab-based two-tier results, correctly differentiating between LD and look-alike diseases. This LD diagnostic test could potentially replace the cumbersome two-tier testing, improving diagnosis and enabling earlier treatment while facilitating immune monitoring and surveillance.

B-109 Outcome of a Large Multicenter Prospective Clinical Study Aimed at Validating a Newly Developed VlsE1/pepC10 IgG/IgM Chemiluminescent Immunoassay (CLIA) in Conjunction with a New Automated Analyzer

Abstract Background Lyme borreliosis is the fastest growing vector-borne illness in North America, with recent estimates as high as 476 000 new cases annually in the U.S. alone. Measurement of Borrelia-specific antibodies via serologic testing remains an important diagnostic aid. Since 1994, serologic testing for Lyme disease in the United States has consisted of a two-stage algorithm, termed standard two-tiered testing (STTT). In the STTT algorithm, specimens equivocal/positive by first tier screening methodologies are subsequently tested via western blots. In recent years, the FDA has cleared several alternatives for replacing western blots with more sensitive second-tier tests, termed modified two-tiered testing (MTTT). We recently developed a VlsE1/pepC10 IgG/IgM chemiluminescent immunoassay (CLIA), to be used in conjunction with a new automated analyzer. The goal of these studies was to conduct a multi-site validation for this new assay and analyzer as a first-tier screening test within an STTT algorithm. Methods Retrospective cohort: 280 clinically characterized samples were provided from the U.S. Centers for Disease Control and Prevention (90 cases, 190 controls). The CDC provided VlsE1/pepC10 IgG/IgM ELISA data derived from their own testing. VlsE1/pepC10 IgG/IgM CLIA and western blotting data were also generated for this panel at ZEUS Scientific. Sensitivity and specificity values were calculated for the retrospective cohort as follows:VlsE1/pepC10-CLIA vs existing VlsE1/pepC10 IgG/IgM-ELISA, and VlsE1/pepC10-CLIA + Western blots, vs VlsE1/pepC10 IgG/IgM-ELISA + Western blots, interpreted as STTT algorithms. All testing of the retrospective cohort was performed at ZEUS Scientific. Prospective Cohort: 600 routine Lyme serology specimens were prospectively collected at three locations (1800 total): Mayo Clinic (MOC), Rochester MN.; Marshfield Clinic (MC), Stevens Point, WI; Health Network Laboratories (HNL), Allentown PA. The samples were assayed and analyzed in the same combinations as described above for the retrospective cohort; however, instead of sensitivity and specificity, the results were reported as positive percent agreement (PPA) and negative percent agreement (NPA) since the prospective samples were not clinically characterized. Testing of the prospective cohort was evenly split across three different laboratories, one of which was ZEUS Scientific. Results Retrospective cohort: [VlsE1/pepC10 CLIA, Sensitivity = (77/90) = 85.56%, Specificity = (177/190) = 93.16%; VlsE1/pepC10 ELISA, Sensitivity = (80/90) = 88.89%, Specificity = (179/190) = 94.21%], [STTT-VlsE1/pepC10-CLIA + Western blots, Sensitivity = (72/90) = 80.00%, Specificity = (190/190) = 100.00%; STTT-VlsE1/pepC10-ELISA + Western blots, Sensitivity = (70/90) = 77.78%, Specificity = (190/190) = 100.00%]. Prospective Cohort (1 sample out of the 1800 collected was omitted due to an invalid western blot result):[VlsE1/pepC10-CLIA vs VlsE1/pepC10-ELISA, PPA = (196/235) = 86.51%, NPA = (1452/1564) = 92.84%; STTT-VlsE1/pepC10-ELISA vs STTT-VlsE1/pepC10-ELISA, PPA = (160/167) = 95.81%, NPA = (1607/11 632) = 98.47%]. Conclusion These studies demonstrate that the newly developed VlsE1/pepC10 IgG/IgM CLIA, in conjunction with a new automated analyzer, yielded accurate results as a first-tier screening assay for both the retrospective and prospective multi-site validation cohorts. Future studies, utilizing the same sample cohorts, will be aimed at validating additional second tier CLIA tests as part of an MTTT algorithm.

Evaluation of the Rapid Quidel Sofia 2 Lyme Immunoassay as a First-Tier Test in a Two-Tier Testing Algorithm for Lyme Disease: Comparison to the Zeus ELISA Borrelia VlsE1/pepC10 IgG/IgM Assay Followed by Immunoblot.

Two-tiered serologic testing for Lyme disease is usually performed using an enzyme-linked immunosorbent assay (ELISA) as the first-tier test. The Quidel Sofia 2 Lyme test is a relatively new lateral flow method to provide more rapid turnaround time. We evaluated its performance in comparison to an established ELISA method. The test can be performed on demand rather than batching assays in a central laboratory. We compared the Sofia 2 assay to the Zeus VlsE1/pepC10 IgG/IgM test in a standard two-tiered testing algorithm. Comparison of the Sofia 2 to the Zeus VlsE1/pepC10 IgG/IgM showed an overall agreement of 89.9% (κ statistic of 0.750, indicating "substantial agreement"). When the tests were followed by immunoblot in a two-tier algorithm, the agreement was 98.9% (κ statistic of 0.973, indicating "almost perfect" agreement). The Sofia 2 Lyme test performs well when compared with the Zeus VlsE1/pepC10 IgG/IgM in a two-tiered testing algorithm.

Lyme Carditis Accompanied with Pseudomonal Pneumonia: A Case Report

Lyme disease is a vastly underdiagnosed disease, and its frequency is steadily rising. It is commonly diagnosed clinically and treated empirically, due to the time required for testing and the inefficiency of laboratory testing methods. Although there are a few reported cases of Lyme carditis, the variation in the clinical presentation and the treatment provided differ significantly. Herein, we present the case of a man who presented in July 2022 with a non-productive cough and low-grade fever along with mild body aches. The patient had an incidental atrial flutter with a 2:1 atrioventricular block, a large thick-walled cavitary lesion in the apex of the left lung, left upper lobe ground-glass opacities, and scattered micronodules in the left lower lobe. He was clinically diagnosed with Lyme carditis despite testing negative for Lyme antibodies. This was in combination with Pseudomonal pneumonia. Here we review recent cases of Lyme carditis and discuss the difficulty of the efficiency of serological testing for Lyme disease.

Maternal Facial Nerve Palsy and a Perinatal Infection.

Maternal Facial Nerve Palsy and a Perinatal Infection.

Lyme disease: "End of the debate?"

Lyme borreliosis is a tick-borne disease that is widespread throughout the northern hemisphere. Ixodes ricinus is present throughout metropolitan France, except for the Mediterranean region. The debate revolves around whether or not a chronic form of Lyme disease exists. This controversy is not limited to France but has been reported worldwide. In France, in 2019, 24 scientific societies representing the medical disciplines most involved in Lyme disease, including the Société Française de Rhumatologie (French Rheumatology Society [SFR]) and the Société de Pathologie Infectieuse de la Langue Française (French Infectious Disease Society-SPILF), published recommendations on the management of Lyme borreliosis following a submission to the Director General of Health. These recommendations conflict with those of the Haute Autorité de Santé (HAS), a multi-specialties independent group of physician, on a key point: whether to add a new nosological entity labeled as "persistent polymorphous signs and symptoms (or syndrome) possibly due to tick bite." The creation of this new syndrome risks should increase anchoring bias, leading to the attribution of all symptoms to a possible tick bite, without considering differential diagnoses. Lyme disease has been extensively studied. Erythema migrans is the primary clinical manifestation. In the presence of nonmetabolic, nonseptic monoarthritis involving the knee or radiculitis of a lower limb during the summer, Lyme disease should be suspected. Serologic testing for Lyme disease is reliable in the case of late forms such as chronic arthritis, while the detection of Borrelia DNA in synovial fluid by PCR is inconsistent. Sometimes, the serology can be misleading in early forms such as radiculitis. Treatment is based on doxycycline for 14 days in early forms (radiculitis), or 28 days in late forms (arthritis). Arthritis can persist or recur after antibiotic therapy. The prevalence of a diffuse polyalgia syndrome (fibromyalgia) following Lyme disease does not seem to differ much from that in the general population. It is not improved by prolonged antibiotic therapy, which is therefore not recommended.

Modified two-tiered testing algorithm for Lyme disease serology: the Canadian context.

Lyme disease (LD) is emerging in many parts of central and eastern Canada. Serological testing is most commonly used to support laboratory diagnosis of LD. Standard two-tiered testing (STTT) for LD involves detection of Borrelia burgdorferi antibodies using an enzyme immunoassay (EIA) followed by IgM and/or IgG immunoblots. However, improved sensitivity has been demonstrated using a modified two-tiered testing (MTTT) approach, in which a second EIA instead of the traditional immunoblot is used. This article summarises the evidence supporting the MTTT versus STTT for laboratory diagnosis of LD in Canada. Peer reviewed literature on the sensitivity and specificity of different EIAs were compared by Canadian experts in LD diagnostic for MTTT vs STTT in patients with clinical history of LD residing in LD endemic areas or in samples from the LD serum repository. The MTTT approach consistently demonstrated improved sensitivity to detect early infections with B. burgdorferi and also maintained high specificity vs STTT. Diagnostic improvements in sensitivity of LD testing without significant loss of specificity have been consistently reported when MTTT is compared with STTT in studies conducted in highly LD endemic regions. Our working group agrees with the recommendation by the United States Centers for Disease Control that serological testing for LD using MTTT is an acceptable alternative to STTT. This recommendation is contingent on development and implementation of comprehensive validation studies on the performance of MTTT vs STTT within the Canadian context, including evaluation of the test performance in areas of low endemicity for LD.

What is the real number of Lyme disease cases in Canada?

BackgroundLyme disease is emerging in Canada due to expansion of the range of the tick vector Ixodes scapularis from the United States. National surveillance for human Lyme disease cases began in Canada in 2009. Reported numbers of cases increased from 144 cases in 2009 to 2025 in 2017. It has been claimed that few (< 10%) Lyme disease cases are reported associated with i) supposed under-diagnosis resulting from perceived inadequacies of serological testing for Lyme disease, ii) expectation that incidence in Canadian provinces and neighbouring US states should be similar, and iii) analysis of serological responses of dogs to the agent of Lyme disease, Borrelia burgdorferi.We argue that performance of serological testing for Lyme disease is well studied, and variations in test performance at different disease stages are accounted for in clinical diagnosis of Lyme disease, and in surveillance case definitions. Extensive surveillance for tick vectors has taken place in Canada providing a clear picture of the emergence of risk in the Canadian environment. This surveillance shows that the geographic scope of I. scapularis populations and Lyme disease risk is limited but increasing in Canada. The reported incidence of Lyme disease in Canada is consistent with this pattern of environmental risk, and the differences in Lyme disease incidence between US states and neighbouring Canadian provinces are consistent with geographic differences in environmental risk. Data on serological responses in dogs from Canada and the US are consistent with known differences in environmental risk, and in numbers of reported Lyme disease cases, between the US and Canada.ConclusionThe high level of consistency in data from human case and tick surveillance, and data on serological responses in dogs, suggests that a high degree of under-reporting in Canada is unlikely. We speculate that approximately one third of cases are reported in regions of emergence of Lyme disease, although prospective studies are needed to fully quantify under-reporting. In the meantime, surveillance continues to identify and track the ongoing emergence of Lyme disease, and the risk to the public, in Canada.

Evaluation of in vivo expressed Borrelia burgdorferi antigens for improved IgM serodiagnosis of early Lyme disease

Improved serologic tests are needed for accurate diagnosis and proper treatment of early stage Lyme disease. We evaluated the 3 antigens currently used for 2-tiered IgM immunoblot testing (FlaB, OspC, and BmpA) in combination with 3 additional antigens (BBA65, BBA70, and BBA73) and measured the sensitivity and specificity against a serum repository of positive and negative controls. Using 3 statistical methods for positivity cutoff determinations and scoring criteria, we found increased sensitivities for early Lyme disease when 2 of 6 antigens were positive as compared with the 2 of 3 antigen IgM criteria currently used for second-tier immunoblot scoring. Specificities for negative controls were comparable or superior to using 2 of 3 antigens. These results indicate that IgM sensitivity and specificity of serological testing for Lyme disease in the early stages of illness can be improved by employing antigens that target the initial host antibody responses.

Clinical presentation of Lyme disease in the higher-risk region of Quebec: a retrospective descriptive study.

Lyme disease is emerging in Canada. This study aimed to describe the use of serologic testing for the disease in the La Pommeraie health region in southern Quebec between 2012 and 2015 and to describe the clinical presentation of laboratory-confirmed cases. The medical charts of all patients investigated for Lyme disease at the Brome-Missisquoi-Perkins Hospital's laboratory between 2012 and 2015 were reviewed for results of serologic testing. Laboratory diagnosis was based on 2-tiered testing: cases had to have positive results of both the enzyme immunoassay and the Western blot test (IgM or IgG). We collected data on clinical presentation for patients assessed at the hospital or at the La Pommeraie Family Medicine Unit. Over the study period, 720 patients were investigated for Lyme disease. There was a more than fivefold increase in requests for serologic testing from 2012 (53) to 2015 (273). The number of confirmed cases increased from 2012 (3) to 2013 (19) and remained stable thereafter (19 in 2014, 18 in 2015). Fifty patients were positive for IgM with or without IgG positivity, and 9 patients were IgG-positive only. Chart reviews were completed for 278 of the 720 patients, including 38 of the 59 laboratory-confirmed cases. Among the 29 IgM-positive patients, the most common symptoms were fever (17 patients [59%]), fatigue (14 [48%]), myalgia (12 [41%]) and headaches (10 [34%]). Twenty-three (79%) had some cutaneous manifestation, including specifically erythema migrans (14 [48%]). A tick bite was reported by 11 patients (38%). Of the 44 patients in the entire study population who presented with erythema migrans, 15 (34%) had confirmed Lyme disease. Requests for serologic testing for Lyme disease increased in the La Pommeraie health region over recent years. Cutaneous manifestations, fever and myalgia were the most common symptoms of IgM-positive cases. Most patients did not report a tick bite.

The Past, Present, and (Possible) Future of Serologic Testing for Lyme Disease

Lyme disease prevails as the most commonly transmitted tick-borne infection in the United States, and serologic evaluation for antibodies to Borrelia burgdorferi remains the recommended modality for diagnosis. This review presents a brief historical perspective on the evolution of serologic assays for Lyme disease and provides a summary of the performance characteristics for the currently recommended two-tiered testing algorithm (TTTA). Additionally, a recently proposed alternative to the traditional TTTA is discussed, and novel methodologies, including immuno-PCR and metabolic profiling for Lyme disease, are outlined.

Secondhand Tobacco Smoke Exposure Among Hospitalized Nonsmokers With Coronary Heart Disease

Secondhand Tobacco Smoke Exposure Among Hospitalized Nonsmokers With Coronary Heart Disease

A concise critical analysis of serologic testing for the diagnosis of lyme disease.

Diagnostic testing for Lyme disease in the clinical setting primarily relies on assessment of serologic responses to infection, with the exception of the early localized phase of disease, in which the diagnosis must be made clinically, due to the recognized insensitivity of serologic testing at this phase of disease. For the diagnosis of early disseminated and late disease, the Centers for Disease Control and Prevention (CDC) recommends a two-tiered approach to testing consisting of initial IgM and IgG quantitative enzyme-linked immunosorbent assay (ELISA), followed by confirmation of all indeterminate or positive ELISA tests with separate IgG and IgM Western blots. This critical analysis addresses the sensitivity, specificity, and predictive value of serologic testing for Lyme disease in early localized, early disseminated, and late disease. Other testing modalities currently under evaluation are also discussed, including IgG vlsE C6 peptide ELISA, other two-tiered testing strategies, rapid diagnostics, and PCR. An understanding of the strengths and limitations of currently available testing for Lyme disease is critical for appropriate diagnosis.

Lyme Disease Associated With Sudden Sensorineural Hearing Loss

Is sudden sensorineural hearing loss associated with Lyme disease in adults? A retrospective case report and a systematic literature search in PubMed and Embase were performed. We describe a patient presenting with sudden sensorineural hearing loss, followed by a facial paralysis, which could be attributed to Lyme disease confirmed by positive serology and a positive immunoblot. She was successfully treated with ceftriaxone, with recovery of the facial paralysis, although no recovery of the hearing loss was observed. A systematic literature search resulted in 4 relevant and valid articles revealing that confirmed positive serology for Borrelia burgdorferi varies from 0% to 21.3%, suggesting active Lyme disease as a cause in patients with sudden sensorineural hearing loss. Two studies demonstrated a significantly higher incidence of confirmed positive serology for Borrelia burgdorferi as compared with the incidence in the general local population. Literature suggests that sudden sensorineural hearing loss may coincide with Borrelia burgdorferi infection. A higher incidence of confirmed positive serology for Borrelia burgdorferi in patients with sudden deafness seems to be depending on the country and on the tests used to confirm Lyme disease. This should be taken into account if serologic testing for Lyme disease in patients with sudden deafness is considered.

Editorial Commentary: Better Laboratory Testing for Lyme Disease: No More Western Blot

It seems unlikely, in routine clinical practice, that direct methods, including culture or polymerase chain reaction, will ever become the primary tests for the laboratory diagnosis of Lyme disease. Indirect, serologic tests will continue to be used to support this diagnosis [1]. Can serologic testing for Lyme disease be improved? The Centers for Disease Control and Prevention (CDC) currently recommends a 2-tiered algorithm for serodiag

High frequency of false positive IgM immunoblots for Borrelia burgdorferi in Clinical Practice

Although it is known that two-tier serologic testing for Lyme disease may be associated with false positive results on the IgM immunoblot, this problem has never been systematically studied in the clinical practice setting. In a retrospective investigation of patients referred to the private adult practice of an Infectious Diseases physician for possible for Lyme disease, 50 of 182 patients (27.5%, 95% CI: 21.1-34.6) were found to have a false positive IgM immunoblot. 78.0% of these patients had received unnecessary antibiotic therapy. False positive results were not restricted to any single commercial laboratory. Research on alternative testing strategies that eliminate the IgM immunoblot entirely is warranted.

Increased Sensitivity and Specificity ofBorrelia burgdorferi16S Ribosomal DNA Detection

The DNA of Borrelia burgdorferi spirochetes extracted by ammonium hydroxide was used as the template for nested polymerase chain reaction (PCR) amplification of the species-specific 16S ribosomal DNA (rDNA). The primers were those well known to be specific for signature sequence amplification of the B burgdorferi sensu lato 16S ribosomal RNA gene. The positive 293-base-pair nested PCR amplicon was subjected to routine direct automated Sanger sequencing. A 50-base sequence excised randomly from the sequencing electrophoretogram between the 2 nested PCR primer binding sites was sufficient for the Basic Local Alignment Search Tool (BLAST) analysis to validate the B burgdorferi sensu lato 16S rDNA without a reasonable doubt. Nested PCR increased the sensitivity of DNA detection by 100- to 1,000-fold. DNA sequence validation based on BLAST algorithms using the GenBank database practically eliminates any possibility of false-positive results due to molecular misidentification. This technology may be a valuable supplement to the current serologic tests for Lyme disease.

Serologic Tests for Lyme Disease: More Smoke and Mirrors

Serologic Tests for Lyme Disease: More Smoke and Mirrors

Prospective Study of Serologic Tests for Lyme Disease

Tests to determine serum antibody levels-the 2-tier sonicate immunoglobulin M (IgM) and immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and Western blot method or the IgG of the variable major protein-like sequence-expressed (VlsE) sixth invariant region (C6) peptide ELISA method-are the major tests available for support of the diagnosis of Lyme disease. However, these tests have not been assessed prospectively. We used these tests prospectively to determine serologic responses in 134 patients with various manifestations of Lyme disease, 89 patients with other illnesses (with or without a history of Lyme disease), and 136 healthy subjects from areas of endemicity and areas in which the infection was not endemic. With 2-tier tests and the C6 peptide ELISA, only approximately one-third of 76 patients with erythema migrans had results that were positive for IgM or IgG seroreactivity with Borrelia burgdorferi in acute-phase samples. During convalescence, 3-4 weeks later, almost two-thirds of patients had seroreactivity with the spirochete B. burgdorferi. The frequencies of seroreactivity were significantly greater among patients with spirochetal dissemination than they were among those who lacked evidence of disseminated disease. Of the 44 patients with Lyme disease who had neurologic, heart, or joint involvement, all had positive C6 peptide ELISA results, 42 had IgG responses with 2-tier tests, and 2 patients with facial palsy had only IgM responses. However, among the control groups, the IgG Western blot was slightly more specific than the C6 peptide ELISA. The differences between the 2 test systems (2-tier testing and C6 peptide ELISA) with respect to sensitivity and specificity were not statistically significant. Except in patients with erythema migrans, both test systems were sensitive for support of the diagnosis of Lyme disease. However, with current methods, 2-tier testing was associated with slightly better specificity.

Is Gulf War Syndrome actually chronic Lyme disease?

Symptoms of Gulf War Syndrome and chronic Lyme disease are very similar. Lyme disease is a condition which can be difficult to diagnose since one of the main features of the condition, the erythema migrans rash, may be absent or overlooked and serological testing for Lyme disease may be falsely negative. Symptoms of Lyme disease may not became apparent until years after exposure to the causative organism. Military personnel during training in the field are at risk of tick bites and it may be that those who developed Gulf War Syndrome entered the conflict with latent Lyme disease. There has been no systematic examination of Gulf War Syndrome sufferers for chronic Lyme disease and it is hypothesized that chronic Lyme disease has been overlooked as a cause of Gulf War Syndrome. To address this it is suggested that sufferers of Gulf War Syndrome or similar illnesses should be examined by physicians who have experience diagnosing and treating large numbers of patients with Lyme disease.