scholarly journals Utility of Bartonella henselae IgM Western Blot Bands for Serodiagnosis of Cat Scratch Disease

2017 ◽  
Vol 56 (1) ◽  
Author(s):  
Ken-ichiro Otsuyama ◽  
Hidehiro Tsuneoka ◽  
Hiroka Yoshidomi ◽  
Mio Haraguchi ◽  
Masashi Yanagihara ◽  
...  
Keyword(s):  
Western Blot ◽  
Bartonella Henselae ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cat Scratch Disease ◽  
Content Type ◽  
Specificity And Sensitivity ◽  
Igm Elisa ◽  
Positive Rate ◽  
Healthy Control

ABSTRACTWe evaluated the utility of Western blot (WB) bands ofBartonella henselae in detecting anti-B. henselaeimmunoglobulin M (IgM) for serodiagnosis of cat scratch disease (CSD). IgM band patterns were examined using sera from 92 patients clinically suspected of having CSD and from 130 healthy individuals. Positive WB bands were observed in 49 (53.5%) of the 92 patient sera. Three bands at 8 to 10, 31 to 35, and 70 kDa were regarded as relevant forB. henselaebecause all of the positive sera yielded at least one of the three bands, and none of the healthy control sera showed reactivity to any of them. In contrast, the positive rate of the patient sera by conventional indirect fluorescence antibody assay (IFA) forB. henselaeIgM was 28.3% (26/92) among the patients. These finding suggest that the IgM-WB assay, although cumbersome to perform, can be used for confirmatory diagnosis of CSD with no false positivity in the control sera. Purification of proteins in the specific bands may contribute to the development of an IgM enzyme-linked immunosorbent assay (IgM-ELISA) with improved specificity and sensitivity.

scholarly journals Development of a Highly Specific IgM Enzyme-Linked Immunosorbent Assay for Bartonella henselae Using RefinedN-Lauroyl-Sarcosine-Insoluble Proteins for Serodiagnosis of Cat Scratch Disease

2016 ◽  
Vol 54 (4) ◽  
pp. 1058-1064 ◽  
Author(s):  
Ken-ichiro Otsuyama ◽  
Hidehiro Tsuneoka ◽  
Kaori Kondou ◽  
Masashi Yanagihara ◽  
Nobuko Tokuda ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Bartonella Henselae ◽  
Fluorescent Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Soluble Antigen ◽  
Cat Scratch Disease ◽  
Healthy Individuals ◽  
Content Type ◽  
Igm Elisa ◽  
Lauroyl Sarcosine

The conventional anti-Bartonella henselaeIgM enzyme-linked immunosorbent assay (IgM-ELISA) methods for diagnosing cat scratch disease (CSD) remain poor in both sensitivity and specificity. We sought to develop an IgM-ELISA with improved accuracy in the serodiagnosis of CSD by exploring the antigens that are most suitable for an ELISA. We prepared 5 different protein antigens: antigen I (sonicatedB. henselaewhole-cell antigen), antigen II (N-lauroyl-sarcosine-insoluble antigen), antigen III (processed sarcosine-soluble antigen), and antigen IV and antigen V (sarcosine-insoluble and sarcosine-soluble antigens refined by DEAE-Sepharose Fast Flow ion-exchange chromatography). The IgM antibodies in the sera of 47 patients with clinically suspected CSD (24 definite, 23 suspected) and of 85 healthy individuals were examined by ELISAs using the 5 antigens, and the results were compared with those of an IgM indirect fluorescent antibody assay (IgM-IFA). In a reference panel, which consisted of 5 positive and 5 negative sera, antigen I and antigen III failed to distinguish between the two statuses, whereas the other three antigens succeeded in distinguishing between them. When the cutoff value was set at the 98th percentile of the ELISA index for healthy individuals, the sensitivity of IgM-IFA for the 24 cases of definite CSD was 54%, whereas the sensitivities of the IgM-ELISAs with antigen II, IV, and V were 75%, 83%, and 75%, respectively. The sensitivities of these three IgM-ELISAs for all 47 of the clinically suspected cases were 49%, 64%, and 51%, respectively. In contrast, the sensitivity of IgM-IFA was 28%. These results indicate that the refined sarcosine-insoluble proteins (antigen IV), which possessed the highest specificity among the 5 antigens, are the most appropriate for developing an IgM-ELISA for the highly specific serodiagnosis of CSD.

scholarly journals Development of an Immunoglobulin M Capture-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Acute Infections with Bartonella henselae

2008 ◽  
Vol 16 (2) ◽  
pp. 282-284 ◽  
Author(s):  
John G. Hoey ◽  
Fernando Valois-Cruz ◽  
Hannah Goldenberg ◽  
Yekaterina Voskoboynik ◽  
Jenna Pfiffner ◽  
...  
Keyword(s):  
Antibody Response ◽  
Immunosorbent Assay ◽  
Bartonella Henselae ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cat Scratch Disease ◽  
Specific Enzyme ◽  
Negative Patient ◽  
Bacillary Angiomatosis ◽  
Acute Infections

ABSTRACT We describe the development of an immunoglobulin M-specific enzyme-linked immunosorbent assay for the detection of an early antibody response to Bartonella henselae, the causative agent of cat scratch disease, bacillary angiomatosis, and endocarditis. This assay discriminates between B. henselae-positive and -negative patient samples with sensitivity and specificity values of 100% and 97.1%, respectively.

scholarly journals Significantly Improved Accuracy of Diagnosis of Early Lyme Disease by Peptide Enzyme-Linked Immunosorbent Assay Based on the Borreliacidal Antibody Epitope of Borrelia burgdorferi OspC

2008 ◽  
Vol 15 (6) ◽  
pp. 981-985 ◽  
Author(s):  
Dean A. Jobe ◽  
Steven D. Lovrich ◽  
Krista E. Asp ◽  
Michelle A. Mathiason ◽  
Stephanie E. Albrecht ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Western Blot ◽  
Immunosorbent Assay ◽  
Antibody Test ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
C Terminus ◽  
Igm Elisa ◽  
Improved Accuracy

ABSTRACT Highly specific borreliacidal antibodies are induced by infection with Borrelia burgdorferi, and the immunodominant response during early Lyme disease is specific for an epitope within the 7 amino acids nearest the C terminus of OspC. We evaluated the ability of an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (OspC7) that matched the region to detect the response and compared the sensitivity during early Lyme disease to that for an FDA-approved Western blot. When the optical density value was adjusted to 98% specificity based on the results from testing normal or uncharacterized sera (n = 236) or sera from patients with blood factors or illnesses that commonly produce antibodies that cross-react with B. burgdorferi antigens (n = 77), 115 (73%) of 157 sera from patients likely to have early Lyme disease were positive for immunoglobulin M (IgM) antibodies and 17 (11%) also had IgG antibodies. In addition, the IgM ELISA reactivities and the titers of antibodies detected by a flow cytometric borreliacidal antibody test correlated closely (r = 0.646). Moreover, the IgM ELISA was significantly more sensitive (P < 0.001) than the Western blot procedure. The findings therefore confirmed that the peptide IgM ELISA detected OspC borreliacidal antibodies and provided strong evidence that the test can eliminate the necessity for confirming early Lyme disease by a supplementary test such as Western blotting.

Serologic Responses to Bartonella and Afipia Antigens in Patients With Cat Scratch Disease

PEDIATRICS ◽  
1995 ◽  
Vol 96 (6) ◽  
pp. 1137-1142
Author(s):  
Cynthia M. Szelc-Kelly ◽  
Simin Goral ◽  
Guillermo I. Perez-Perez ◽  
Bradley A. Perkins ◽  
Russell L. Regnery ◽  
...  
Keyword(s):  
Immunoglobulin M ◽  
Skin Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Cat Scratch Disease ◽  
Control Subjects ◽  
Number Of Patients ◽  
Healthy Control ◽  
Antibody Levels ◽  
And Control

Objective. To assess the serologic response to Afipia and Bartonella, previously named Rochalimaea, in patients with cat scratch disease (CSD) and a healthy control group. Design. Prospective, controlled trial. Setting. Referral clinic and hospitalized patients in a university medical center. Participants. Eighty patients with CSD and 57 healthy control subjects of similar age. Main Outcome Measures. The immune responses to Afipia felis and Bartonella henselae were evaluated by a newly developed enzyme-linked immunosorbent assay (ELISA) in patients with CSD and healthy control subjects. Responses to B henselae were also measured by indirect fluorescent antibody (IFA) tests. Antibody levels to Bartonella quintana were measured by ELISA and IFA in a limited number of patients and control subjects. Results. Of the 80 patients with clinical CSD, 56 had positive results of CSD skin tests. ELISA antibody levels to A felis did not differ between patients and control subjects, but immunoglobulin M (IgM) and IgG ELISA antibodies to B henselae and B quintana were significantly higher in patients than in control subjects. IFA responses to B henselae and B quintana were also significantly higher in patients than in control subjects. Conclusion. Patients with CSD had significant serologic responses to B henselae and B quintana but not to A felis, suggesting that the causative agent of CSD is antigenically related to the Bartonella genus and not to Afipia. The Bartonella IgM ELISA and IFA assay were both sensitive and specific and may be used to establish the diagnosis of CSD.

scholarly journals Cervical Cat Scratch Disease Lymphadenitis in a Patient with Immunoglobulin M Antibodies to Toxoplasma gondii

2002 ◽  
Vol 9 (2) ◽  
pp. 496-498
Author(s):  
Mardjan Arvand ◽  
Ilkay Kazak ◽  
Sergije Jovanovic ◽  
Hans-Dieter Foss ◽  
Oliver Liesenfeld
Keyword(s):  
Toxoplasma Gondii ◽  
Clinical Symptoms ◽  
Bartonella Henselae ◽  
Cervical Lymphadenopathy ◽  
Immunoglobulin M ◽  
Cat Scratch Disease ◽  
Specific Immunoglobulin ◽  
Acute Toxoplasmosis

ABSTRACT We report on a young patient with chronic cervical lymphadenopathy and serological and histological evidence for infection with Bartonella henselae and Toxoplasma gondii. Serological follow-up studies, including testing for avidity of Toxoplasma-specific immunoglobulin G antibodies, assisted in the determination of the cause of the acute lymphadenitis. Our results suggest that the clinical symptoms were most likely due to cat scratch disease rather than to acute toxoplasmosis.

scholarly journals Development of a Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycoplasma bovis Infection in Cattle

2013 ◽  
Vol 21 (2) ◽  
pp. 196-202 ◽  
Author(s):  
Nadeeka K. Wawegama ◽  
Glenn F. Browning ◽  
Anna Kanci ◽  
Marc S. Marenda ◽  
Philip F. Markham
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Mycoplasma Bovis ◽  
Western Blots ◽  
Content Type ◽  
Amino Terminal ◽  
Igm Elisa ◽  
Specific Igg ◽  
Lipase A ◽  
Strong Immunoreactivity ◽  
Gst Fusion Proteins

ABSTRACTMycoplasma boviscauses a range of diseases in cattle, including mastitis, arthritis, and pneumonia. However, accurate serological diagnosis of infection remains problematic. The studies described here aimed to identify an antigen that might be used to develop a more specific and sensitive diagnostic assay. A 226-kDa immunogenic protein was consistently detected in Western blots by antibodies in sera from calves experimentally infected withM. bovis. This protein was shown to be a membrane protein with lipase activity and was named mycoplasma immunogenic lipase A (MilA). Different regions of MilA were expressed inEscherichia colias glutathioneS-transferase (GST) fusion proteins and recombinant products from the amino-terminal end shown to have strong immunoreactivity withM. bovis-specific bovine sera. The most immunoreactive fusion protein, GST-MilA-ab, was used to develop indirect IgM and IgG enzyme-linked immunosorbent assays (ELISAs). The IgM ELISA detectedM. bovis-specific IgM antibody 2 weeks after infection with 97.1% sensitivity and had a specificity of 63.3%, while the IgG ELISA detectedM. bovis-specific IgG 3 weeks after infection with 92.86% sensitivity and had a specificity of 98.7%, demonstrating that the IgG ELISA has potential for use as a sensitive and specific assay for detecting infection in cattle.

scholarly journals Comparison of Western Immunobloting to an Enzyme-Linked Immunosorbent Assay for the Determination of Anti-Bordetella pertussis Antibodies

2011 ◽  
Vol 18 (4) ◽  
pp. 615-620 ◽  
Author(s):  
Stephen D. Merrigan ◽  
Ryan J. Welch ◽  
Christine M. Litwin
Keyword(s):  
Immunosorbent Assay ◽  
Bordetella Pertussis ◽  
World Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Infection ◽  
Content Type ◽  
Igm Elisa ◽  
Iga Antibodies ◽  
Health Organization

ABSTRACTDuringBordetella pertussisinfection, it has been established that an increase of anti-pertussis toxin (PT) and anti-filamentous hemagglutinin (FHA) antibodies occurs. Immunoblots from two manufacturers using FHA and PT antigens were compared with an enzyme-linked immunosorbent assay (ELISA) that used both FHA and PT. One manufacturer used two concentrations of PT bands for the IgG immunoblot, calibrated to the World Health Organization standard for PT in international units (IU/ml), 100 IU/ml (PT-100) and 8 IU/ml (PT). The second immunoblot kit measured antibodies to a single calibrated PT band. Both kits measured IgA antibodies, and one additionally measured IgM antibodies. Two of 41 (5%) ELISA IgM positives were confirmed positive by IgM immunoblotting, suggesting poor specificity of the IgM ELISA. The agreements of the IgG and IgA immunoblots with the ELISA ranged from 72.5% to 85.3%, with only 38 to 51% of IgA positives confirmed by immunoblotting and only 61 to 68% of IgG positives confirmed by immunoblotting. The two immunoblots correlated well with each other, with 91.7% and 94.3% agreement for IgG and IgA, respectively. When the FHA band was used with the PT band as the criterion for positivity, significant differences existed in specificity compared to the ELISA (IgG, 84.1% versus 33.3%; IgA, 82.4% versus 71.0%). When the positive IgA immunoblots (evidence of natural recent infection) were compared to the positive PT-100 IgG immunoblots (evidence of recent infection or vaccination), the PT-100 blot showed a 71% sensitivity in detecting natural recent infection.B. pertussisimmunoblots, alone or in combination with ELISAs, can aid in the diagnosis ofB. pertussisinfection.

scholarly journals Development and Evaluation of an Epstein-Barr Virus (EBV) Immunoglobulin M Enzyme-Linked Immunosorbent Assay Based on the 18-Kilodalton Matrix Protein for Diagnosis of Primary EBV Infection

1998 ◽  
Vol 36 (11) ◽  
pp. 3359-3361 ◽  
Author(s):  
K. H. Chan ◽  
R. X. Luo ◽  
H. L. Chen ◽  
M. H. Ng ◽  
W. H. Seto ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Epstein Barr Virus ◽  
Matrix Protein ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Clinical Specimens ◽  
Barr Virus ◽  
Ebv Infection ◽  
Igm Elisa ◽  
Epstein Barr

A new immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) based on the recombinant Epstein-Barr virus (EBV) matrix protein was developed. Compared to indirect immunofluorescence for the detection of IgM antibody to the EBV capsid antigen on clinical specimens, the sensitivity and specificity of the new IgM ELISA were 96 and 96%, respectively.

scholarly journals Performance Characteristics of an In-House Assay System Used To Detect West Nile Virus (WNV)-Specific Immunoglobulin M during the 2001 WNV Season in the United States

2003 ◽  
Vol 10 (1) ◽  
pp. 177-179 ◽  
Author(s):  
Harry E. Prince ◽  
Wayne R. Hogrefe
Keyword(s):  
West Nile Virus ◽  
The United States ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Screening Assay ◽  
West Nile ◽  
Elisa System ◽  
Igm Elisa ◽  
Specific Immunoglobulin ◽  
Control And Prevention

ABSTRACT During the 2001 U. S. West Nile virus (WNV) season, 163 specimens were reactive in an in-house WNV-specific immunoglobulin M (IgM) screening enzyme-linked immunosorbent assay (ELISA) and were referred to either the Centers for Disease Control and Prevention or the appropriate state public health laboratory (CDC/SPHL) for additional testing. CDC/SPHL supplied results for 124 specimens that could be further evaluated in-house: 70 specimens were nonreactive in the CDC/SPHL WNV-specific IgM screening assay, and 54 specimens were reactive. These specimens were used to evaluate a modified in-house WNV-specific IgM ELISA that incorporated background subtraction to identify nonspecific reactivity and thus improve assay specificity. Of the 70 CDC/SPHL nonreactive samples, 49 (70%) were nonreactive in the modified ELISA; of the 54 CDC/SPHL reactive samples, 51 (94%) were reactive in the modified ELISA. Confirmatory studies performed by CDC/SPHL indicated that 38 CDC/SPHL screen-reactive specimens represented true WNV infection; all 38 specimens were reactive in the modified in-house WNV-specific IgM ELISA. These findings demonstrate that an in-house ELISA system for WNV-specific IgM effectively identifies patients with WNV infection.

scholarly journals Two Methods for Rapid Serological Diagnosis of Acute Leptospirosis

2001 ◽  
Vol 8 (2) ◽  
pp. 349-351 ◽  
Author(s):  
Paul N. Levett ◽  
Songee L. Branch ◽  
Carol U. Whittington ◽  
Charles N. Edwards ◽  
Helene Paxton
Keyword(s):  
Positive Predictive Value ◽  
Negative Predictive Value ◽  
Predictive Value ◽  
Immunoglobulin M ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rapid Assays ◽  
Igm Elisa ◽  
Highly Sensitive ◽  
Specialized Equipment

ABSTRACT Leptospirosis is a common and underdiagnosed zoonosis. Two rapid assays for serological diagnosis of acute leptospirosis in diagnostic laboratories, the immunoglobulin M (IgM)-dipstick assay and the indirect hemagglutination assay (IHA), were evaluated and compared with standard assays. Sera were examined from 104 patients admitted to a hospital for investigation in a leptospirosis diagnostic protocol. Specimens for serology were taken on days 1 and 4 of the patients' hospital stay. Antibodies were detected using an IgM-enzyme-linked immunosorbent assay (ELISA), microscopic agglutination test (MAT), an IgM-dipstick assay, and an IHA. Fifty-one patients were found to have leptospirosis. The sensitivity of the IgM-dipstick assay was 98%, its specificity was 90.6%, its positive predictive value was 90.9%, and its negative predictive value was 98%. The sensitivity of the IHA was 92.2%, its specificity was 94.4%, its positive predictive value was 95.9%, and its negative predictive value was 92.7%. The standard IgM-ELISA and MAT, were positive in the first samples tested from 67 and 55% of the cases, respectively, and the rapid IgM-dipstick assay and IHA were positive in 71 and 49%, respectively, in the first sample tested. Both rapid assays are highly sensitive and specific. Neither requires specialized equipment, and both are suitable for use in diagnostic laboratories.

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