scholarly journals C-Terminal Invariable Domain of VlsE Is Immunodominant but Its Antigenicity Is Scarcely Conserved among Strains of Lyme Disease Spirochetes

2001 ◽  
Vol 69 (5) ◽  
pp. 3224-3231 ◽  
Author(s):  
Fang Ting Liang ◽  
Lisa C. Bowers ◽  
Mario T. Philipp
Keyword(s):  
Lyme Disease ◽  
Antibody Response ◽  
Human Serum ◽  
Synthetic Peptide ◽  
Species Complex ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Variable Surface ◽  
Entire Sequence ◽  
Carboxyl Terminal

ABSTRACT VlsE, the variable surface antigen of Borrelia burgdorferi, contains two invariable domains located at the amino and carboxyl terminal ends, respectively, and a central variable domain. In this study, both immunogenicity and antigenic conservation of the C-terminal invariable domain were assessed. Mouse antiserum to a 51-mer synthetic peptide (Ct) which reproduced the entire sequence of the C-terminal invariable domain of VlsE from B. burgdorferi strain B31 was reacted on immunoblots with whole-cell lysates extracted from spirochetes of 12 strains from the B. burgdorferi sensu lato species complex. The antiserum recognized only VlsE from strain B31, indicating that epitopes of this domain differed among these strains. When Ct was used as enzyme-linked immunosorbent assay (ELISA) antigen, all of the seven monkeys and six mice that were infected with B31 spirochetes produced a strong antibody response to this peptide, indicating that the C-terminal invariable domain is immunodominant. None of 12 monkeys and only 11 of 26 mice that were infected with strains other than B31 produced a detectable anti-Ct response, indicating a limited antigenic conservation of this domain among these strains. Twenty-six of 33 dogs that were experimentally infected by tick inoculation were positive by the Ct ELISA, while only 5 of 18 serum samples from dogs clinically diagnosed with Lyme disease contained detectable anti-Ct antibody. Fifty-seven of 64 serum specimens that were collected from American patients with Lyme disease were positive by the Ct ELISA, while only 12 of 21 European samples contained detectable anti-Ct antibody. In contrast, antibody to the more conserved invariable region IR6 of VlsE was present in all of these dog and human serum samples.

scholarly journals Sensitive and Specific Serodiagnosis of Lyme Disease by Enzyme-Linked Immunosorbent Assay with a Peptide Based on an Immunodominant Conserved Region of Borrelia burgdorferi VlsE

1999 ◽  
Vol 37 (12) ◽  
pp. 3990-3996 ◽  
Author(s):  
Fang Ting Liang ◽  
Allen C. Steere ◽  
Adriana R. Marques ◽  
Barbara J. B. Johnson ◽  
James N. Miller ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Immunosorbent Assay ◽  
Synthetic Peptide ◽  
Late Phase ◽  
Hospitalized Patients ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Variable Surface ◽  
Conserved Region

VlsE, the variable surface antigen of Borrelia burgdorferi, contains an immunodominant conserved region named IR6. In the present study, the diagnostic performance of a peptide enzyme-linked immunosorbent assay (ELISA) based on a 26-mer synthetic peptide (C6) with the IR6 sequence was explored. Sensitivity was assessed with serum samples (n = 210) collected from patients with clinically defined Lyme disease at the acute (early localized or early disseminated disease), convalescent, or late disease phase. The sensitivities for acute-, convalescent-, and late-phase specimens were 74% (29 of 39), 85 to 90% (34 of 40 to 35 of 39), and 100% (59 of 59), respectively. Serum specimens from early neuroborreliosis patients were 95% positive (19 of 20), and those from an additional group of patients with posttreatment Lyme disease syndrome yielded a sensitivity of 62% (8 of 13). To assess the specificity of the peptide ELISA, 77 serum samples from patients with other spirochetal or chronic infections, autoimmune diseases, or neurologic diseases and 99 serum specimens from hospitalized patients in an area where Lyme disease is not endemic were examined. Only two potential false positives from the hospitalized patients were found, and the overall specificity was 99% (174 of 176). Precision, which was assessed with a panel of positive and negative serum specimens arranged in blinded duplicates, was 100%. Four serum samples with very high anti-OspA antibody titers obtained from four monkeys given the OspA vaccine did not react with the C6 peptide. This simple, sensitive, specific, and precise ELISA may contribute to alleviate some of the remaining problems in Lyme disease serodiagnosis. Because of its synthetic peptide base, it will be inexpensive to manufacture. It also will be applicable to serum specimens from OspA-vaccinated subjects.

scholarly journals Characterization of a Borrelia burgdorferi VlsE Invariable Region Useful in Canine Lyme Disease Serodiagnosis by Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 38 (11) ◽  
pp. 4160-4166 ◽  
Author(s):  
Fang Ting Liang ◽  
Richard H. Jacobson ◽  
Reinhard K. Straubinger ◽  
Amy Grooters ◽  
Mario T. Philipp
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Antibody Response ◽  
Immunosorbent Assay ◽  
Protein A ◽  
Immunoblot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Entire Study ◽  
Variable Surface

Sera collected from dogs experimentally infected withBorrelia burgdorferi by tick inoculation were analyzed for an antibody response to each of the six invariable regions (IRs; i.e., IR1 to IR6) of VlsE, the variable surface antigen of B. burgdorferi. Six synthetic peptides (C1 to C6), which reproduced the six IR sequences were used as peptide-based, enzyme-linked immunosorbent assay (ELISA) antigens. Two IRs, IR2 and IR6, were found to be immunodominant. Studies with serially collected serum samples from experimentally infected dogs revealed that the antibody response to IR6 appears earlier and is stronger than that to IR2. Thus, the IR6 sequence alone appeared to be sufficient for serodiagnosis. When C6 alone was used as antigen, the peptide-based ELISA was positive in 7 of 23 dogs (30%) as early as 3 weeks postinfection. All dogs (n = 33) became strongly positive 1 or 2 weeks later, and this response persisted for the entire study, which lasted for 69 weeks. Of 55 sera submitted by veterinarians from dogs suspected of having Lyme disease, 19 were also positive by the C6 ELISA, compared to 20 positives detected by immunoblot analysis using cultured B. burgdorferi lysates as antigen. The sensitivity of using C2 and C6 together for detecting specific antibody in both experimentally infected and clinically diagnosed dogs was not better than sensitivity with C6 alone, confirming that C6 suffices as a diagnostic probe. Moreover, the C6 ELISA yielded 100% specificity with serum samples collected from 70 healthy dogs, 14 dogs with infections other than B. burgdorferi, and 15 animals vaccinated with either outer surface protein A, whole-spirochete vaccines, or the common puppy-vaccines. Therefore, this C6 ELISA was both sensitive and specific for the serodiagnosis of canine Lyme disease and could be used with vaccinated dogs.

scholarly journals Use of Moraxella catarrhalis Lipooligosaccharide Mutants To Identify Specific Oligosaccharide Epitopes Recognized by Human Serum Antibodies

2009 ◽  
Vol 77 (10) ◽  
pp. 4548-4558 ◽  
Author(s):  
Johanna M. Schwingel ◽  
Katie J. Edwards ◽  
Andrew D. Cox ◽  
Hussein Masoud ◽  
James C. Richards ◽  
...  
Keyword(s):  
Immune Response ◽  
Antibody Response ◽  
Human Serum ◽  
Moraxella Catarrhalis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Antibody ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Carbohydrate Epitopes ◽  
Tract Infections

ABSTRACT Moraxella catarrhalis is a causative agent of otitis media in children and lower respiratory tract infections in adults suffering from chronic obstructive pulmonary disease (COPD). This strict human pathogen continues to be a significant cause of disease in this broad spectrum of patients because there is no available vaccine. Although numerous putative vaccine antigens have been described, little is known about the human immune response to M. catarrhalis infection in vivo. Human serum antibodies are directed at a number of surface proteins, and lipooligosaccharides (LOS) and detoxified LOS may be an effective immunogen in mice. In this study, we used a specific LOS-based enzyme-linked immunosorbent assay (ELISA), containing the three major M. catarrhalis serotypes together with a complete series of truncated LOS mutants, to detect the development of new antibodies to specific regions of the oligosaccharide molecule. We compared serum samples from COPD patients who had recently cleared an M. catarrhalis infection to serum samples collected prior to their infection. Variability in the antibody response to LOS was observed, as some patients developed serotype-specific antibodies, others developed antibodies to the LOS of each serotype, others developed broadly cross-reactive antibodies, and some did not develop new antibodies. These newly developed human antibodies are directed at both side chains and core structures in the LOS molecule. This LOS-based ELISA can be used to dissect the human antibody response to both internal and external carbohydrate epitopes, thus providing a better understanding of the humoral immune response to M. catarrhalis LOS epitopes developed during natural infection.

scholarly journals Evaluation of the Recombinant VlsE-Based Liaison Chemiluminescence Immunoassay for Detection of Borrelia burgdorferi and Diagnosis of Lyme Disease

2008 ◽  
Vol 15 (12) ◽  
pp. 1796-1804 ◽  
Author(s):  
Thomas B. Ledue ◽  
Marilyn F. Collins ◽  
John Young ◽  
Martin E. Schriefer
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Viral Infections ◽  
Relative Sensitivity ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Variable Surface ◽  
Relative Specificity

ABSTRACT Recent efforts to improve the serologic diagnosis of Lyme disease have included the use of a synthetic peptide (C6) that reproduces the sequence of invariable region 6 of VlsE, the variable surface antigen of Borrelia burgdorferi. In the present study, the diagnostic performance of DiaSorin's recombinant VlsE-based chemiluminescence immunoassay in 1,947 human serum samples was evaluated. Sensitivity was determined using two serum panels from the CDC. For panel I, we observed sensitivities of 68.4% and 75.6% for subjects with early, localized (n = 19) or disseminated (n = 41) disease, respectively. For panel II, we observed sensitivities of 61.5% and 100% for subjects with early (n = 26) or late-stage (n = 11) disease, respectively. We observed a specificity of 99.5% for healthy donors (n = 600) living either in regions of the United States where the disease is endemic or in regions where it is not endemic. Overall, specificity among 207 potentially cross-reactive sera from subjects who had other spirochetal infections, nonspirochetal infections including bacterial and viral infections, or autoimmune or neurologic disease; who were positive for rheumatoid factor or anti-mouse antibodies; or who had been previously vaccinated for Lyme disease was 93.7%. In a direct comparison of 1,038 prospectively collected samples for Lyme disease testing we observed a relative sensitivity of 70%, a relative specificity of 99.1%, and an overall agreement of 97.1% between the DiaSorin recombinant VlsE chemiluminescence immunoassay and the Immunetics peptide-based C6 enzyme-linked immunosorbent assay.

scholarly journals Quantitative Approach for the Serodiagnosis of Canine Lyme Disease by the Immunoblot Procedure

2000 ◽  
Vol 38 (7) ◽  
pp. 2628-2632 ◽  
Author(s):  
Marta A. Guerra ◽  
Edward D. Walker ◽  
Uriel Kitron
Keyword(s):  
Logistic Regression ◽  
Lyme Disease ◽  
Natural Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Immunoblot Assay ◽  
Serological Status ◽  
Low Probability ◽  
Positive Results ◽  
Logistic Regression Equation

Serum samples obtained from healthy, asymptomatic dogs in areas of Wisconsin and northern Illinois where Lyme disease is endemic or nonendemic were assayed for antibodies to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), and positive results were confirmed by immunoblot assay. We found that 56.9% (562 of 1,077) of the samples were positive by ELISA and 82.0% (461 of 562) were positive by immunoblotting. A logistic regression model was developed to distinguish between nonvaccinated dogs naturally infected with B. burgdorferi from areas where the disease is endemic and dogs from areas where the disease is nonendemic that were vaccinated against Lyme disease. Of the 18 protein bands analyzed, 8 were found to be significantly different (P < 0.05) between the two groups. p93, p34, p31, and p28 occurred with increased frequency in vaccinated dogs, while p58, p37, p35, and p30 occurred more frequently in naturally infected dogs. The logistic regression equation obtained was used to determine the probability of natural infection among vaccinated dogs residing in areas where the disease is endemic. Of 125 samples, 87.2% had a very low probability of natural infection and only 2.4% were highly likely to be infected. Logistic regression is a useful method for distinguishing between vaccinated and naturally infected dogs and predicting the serological status of vaccinated dogs from areas where Lyme disease is endemic.

scholarly journals Seroprevalence of Jamestown Canyon virus in the Japanese general population

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Hirofumi Kato ◽  
Masaaki Satoh ◽  
Madoka Kawahara ◽  
Satoshi Kitaura ◽  
Tomoki Yoshikawa ◽  
...  
Keyword(s):  
General Population ◽  
Human Serum ◽  
Health Agency ◽  
Febrile Illness ◽  
Central Nervous System Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Assay ◽  
Serum Samples ◽  
Human Serum Samples ◽  
Jamestown Canyon Virus

Abstract Background Jamestown Canyon virus (JCV) is a mosquito-borne orthobunyavirus that causes acute febrile illness, meningitis, and meningoencephalitis, mainly among adults. JCV is widely distributed in North America and the number of JCV cases in the U.S. has increased in recent years. Therefore, the central nervous system disease caused by JCV can be considered a potentially re-emerging viral disease. However, the seroprevalence of JCV is unknown in Japan. The purpose of this study is to evaluate the seroprevalence of JCV in the Japanese population. Methods We used an IgG enzyme-linked immunosorbent assay (IgG-ELISA) with JCV-infected cell-lysates and/or a neutralizing (NT) antibody assay. The cut-off value of IgG-ELISA was determined using IgG-ELISA to analyze serum specimens from 37 healthy Japanese donors. IgG-ELISA was validated by assessing its sensitivity and specificity, using 38 human serum samples previously tested for the presence or absence of antibodies against JCV and snowshoe hare virus (SSHV), in an in-house NT antibody assay conducted by the Public Health Agency of Canada. The seroepidemiological study was performed using IgG-ELISA and NT antibody assay to analyze 246 human serum samples from the serum bank of the National Institute of Infectious Diseases (NIID) in Japan. Results The cut-off value of IgG-ELISA was determined at 0.20, based on the mean (− 0.075) and standard deviation (0.092) values using Japanese donors’ sera. The sensitivity and the specificity of IgG-ELISA determined using 25 JCV-positive and 4 JCV-negative serum samples were 96 and 100%, respectively. Analysis of the 246 Japanese serum samples revealed that no specimen showed a higher value than the cut-off value of IgG-ELISA, and no sample tested positive by the NT antibody assay. Conclusions Our results showed that JCV is not circulating significantly in Japan. To the best of our knowledge, this is the first report to demonstrate the seroprevalence of JCV in the general population in Japan.

DETECTION OF SPECIFIC ANTIBODIES TO THE NUCLEOCAPSID PROTEIN FRAGMENTS OF SEVERE ACUTE RESPIRATORY SYNDROME-CORONAVIRUS AND SCOTOPHILUS BAT CORONAVIRUS-512 IN THREE INSECTIVOROUS BAT SPECIES

2018 ◽  
Vol 44 (04) ◽  
pp. 179-188 ◽  
Author(s):  
Yi-Ning Chen ◽  
Bo-Gang Su ◽  
Hung-Chang Chen ◽  
Cheng-Han Chou ◽  
Hsi-Chi Cheng
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Nucleocapsid Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Protein Fragments ◽  
Specific Antibodies ◽  
History Of ◽  
Polymerase Chain ◽  
Carboxyl Terminal ◽  
Bat Coronavirus

Bats are the natural reservoirs of severe acute respiratory syndrome-coronavirus (SARS-CoV). Six Alphacoronavirus and five Betacoronavirus have been detected in many bat species, including SARS-related CoV and Middle East respiratory syndrome (MERS)-related CoV. In Taiwan, SARS-related CoV, belonging to Betacoronavirus, has been detected in Rhinolophus monoceros. Scotophilus bat CoV-512, belonging to Alphacoronavirus, has been detected in Scotophilus kuhlii, Miniopterus fuliginosus, and Rhinolophus monoceros by using reverse transcription polymerase chain reaction (RT-PCR). To understand the infection history of CoV in these three insectivorous bat populations, CoV-specific antibodies were surveyed by using western blot (WB) analysis and indirect enzyme-linked immunosorbent assay (ELISA). The carboxyl terminal fragment of nucleocapsid protein (N3) of SARS-CoV and Scotophilus bat CoV-512 were used as the antigen in the assays. Of the 52 serum samples obtained from Scotophilus kuhlii, 29 samples (56%) were tested positive for Scotophilus bat CoV-512-specific antibodies through ELISA. Of the 63 serum samples obtained from Rhinolophus monoceros, 9 samples were tested positive for only SARS-CoV-specific antibodies, 7 samples were tested positive for only Scotophilus bat CoV-512-specific antibodies, and 16 samples (25.4%) were tested positive for both antibodies through WB analysis. Only 1 of 18 Miniopterus bat serum samples tested positive for Scotophilus bat CoV-512-specific antibodies through ELISA. Lactating female bats had higher positive rates of CoV-specific antibodies than non-lactating female and male bats did. Our findings were crucial for understanding CoV infection history in three insectivorous bat species and important for the control of bat-borne zoonosis diseases.

scholarly journals Outer Surface Protein C Peptide Derived from Borrelia burgdorferi Sensu Stricto as a Target for Serodiagnosis of Early Lyme Disease

2013 ◽  
Vol 20 (4) ◽  
pp. 474-481 ◽  
Author(s):  
Paul M. Arnaboldi ◽  
Rudra Seedarnee ◽  
Mariya Sambir ◽  
Steven M. Callister ◽  
Josephine A. Imparato ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Bacterial Species ◽  
Erythema Migrans ◽  
Early Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Attractive Alternative ◽  
Serum Samples ◽  
Peptide Epitope ◽  
Content Type

ABSTRACTCurrent serodiagnostic assays for Lyme disease are inadequate at detecting early infection due to poor sensitivity and nonspecificity that arise from the use of whole bacteria or bacterial proteins as assay targets; both targets contain epitopes that are cross-reactive with epitopes found in antigens of other bacterial species. Tests utilizing peptides that contain individual epitopes highly specific forBorrelia burgdorferias diagnostic targets are an attractive alternative to current assays. Using an overlapping peptide library, we mapped linear epitopes in OspC, a critical virulence factor ofB. burgdorferirequired for mammalian infection, and confirmed the results by enzyme-linked immunosorbent assay (ELISA). We identified a highly conserved 20-amino-acid peptide epitope, OspC1. Via ELISA, OspC1 detected specific IgM and/or IgG in 60 of 98 serum samples (62.1%) obtained from patients with erythema migrans (early Lyme disease) at the time of their initial presentation. By comparison, the commercially available OspC peptide PepC10 detected antibody in only 48 of 98 serum samples (49.0%). In addition, OspC1 generated fewer false-positive results among negative healthy and diseased (rheumatoid arthritis and positive Rapid Plasma Reagin [RPR+] test result) control populations than did PepC10. Both highly specific and more sensitive than currently available OspC peptides, OspC1 could have value as a component of a multipeptide Lyme disease serological assay with significantly improved capabilities for the diagnosis of early infection.

scholarly journals Rapid Decline of OspC Borreliacidal Antibodies following Treatment of Patients with Early Lyme Disease

2011 ◽  
Vol 18 (6) ◽  
pp. 1034-1037 ◽  
Author(s):  
Dean A. Jobe ◽  
Todd J. Kowalski ◽  
Marissa Bloemke ◽  
S. D. Lovrich ◽  
Steven M. Callister
Keyword(s):  
Lyme Disease ◽  
Immunosorbent Assay ◽  
Strong Evidence ◽  
Significant Decline ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rapid Decline ◽  
Seropositive Patient ◽  
Serum Samples ◽  
Igm Elisa ◽  
Alternative Test

ABSTRACTWe determined whether the levels of OspC borreliacidal antibodies declined following treatment of early Lyme disease and whether the OspC7 peptide enzyme-linked immunosorbent assay (ELISA) could be used as an alternative test for detecting the response. Serum samples were collected from 37 subjects at the onset of illness and 2 and 6 months after treatment with doxycycline. The ELISA detected IgM and IgG OspC7 antibodies within 2 months in 18 (49%) and 5 (14%) sera, respectively. Moreover, the sera from 12 subjects who tested positive by the ELISA also showed borreliacidal activity which was completely abrogated when the antibodies to OspC7 were removed. The borreliacidal activity decreased greater than 4-fold in each seropositive patient within 6 months after treatment, and the findings were accurately predicted by the IgM ELISA. The results confirmed that the ELISA was an effective alternative for detection of OspC borreliacidal antibodies produced during early Lyme disease in humans and also provided strong evidence that a significant decline in the response coincides with successful treatment of the illness.

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