Development of a Highly Specific IgM Enzyme-Linked Immunosorbent Assay for Bartonella henselae Using Refined
            <i>N</i>
            -Lauroyl-Sarcosine-Insoluble Proteins for Serodiagnosis of Cat Scratch Disease

Otsuyama, Ken-ichiro; Tsuneoka, Hidehiro; Kondou, Kaori; Yanagihara, Masashi; Tokuda, Nobuko; Shirasawa, Bungo; Ichihara, Kiyoshi

doi:10.1128/jcm.03009-15

Cited by 22 publications

(12 citation statements)

References 18 publications

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“…Whole-cell lysate suspensions of B. henselae ATCC 49882 were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and WB analysis. These procedures are the same as those reported previously but with minor modifications (13). Briefly, the B. henselae suspension was separated by SDS-PAGE using a 5-to-20%-gradient polyacrylamide gel (ATTO, Japan).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2018

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We evaluated the utility of Western blot (WB) bands of e in detecting anti- immunoglobulin 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 for because 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) for IgM 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.

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Section: Methodsmentioning

confidence: 99%

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

et al. 2018

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“…So, bartonelloses are often diagnosed through serological tests. However, these tests can be negative since the conventional anti- Bartonella henselae IgM ELISA methods are poor for as far as both sensitivity and specificity are concerned [ 7 ]. In line with data showing that serology is affected by false negative results, among our patients, only 1 out of 3 had positive IgM, while Bartonella DNA was positive in all cases.…”

Section: Discussionmentioning

confidence: 99%

Utility of Molecular Identification and Quantitation of Bartonella Species with Species-Specific Real-Time PCR for Monitoring Treatment Response: A Case Series

Mazzitelli¹,

Lamberti²,

Quirino³

et al. 2018

TOMICROJ

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Background:Bartonella species are intracellular bacteria capable of producing several diseases in humans. The three most common and wellknown diseases are cat scratch disease (CSD), caused by B. henselae, trench fever, caused by B. quintana and Carrion’s Disease, caused by B. bacilliformis. Signs and symptoms are very different and aspecific: Fatigue, fever, headache, lymphadenopathy, malaise, loss of weight. No data exist to support guidelines’ recommendations to decide which drugs should be optimally used and how long they should be administered. Therefore, a marker of treatment response is needed to guide treatment strategies.Methods:We report herein three cases in which a species specific Reverse-Transcriptase Polymerase-Chain-Reaction (RT PCR) developed in-house was performed and compared to serology in order to make diagnosis and to evaluate treatment response.Results:Our species-specific RT PCR seemed to play a fundamental role both in diagnosis and treatment. Moreover, a discrepancy with the serology results was found.Conclusion:Further studies are necessary to validate these results and elucidate what is the best treatment for this pleomorphic disease. However, in absence of clear guidelines, RT PCR may be useful to orientate kind of treatment ad its duration.

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“…Cat scratch disease (CSD) is an emerging infectious disease worldwide caused by Bartonella henselae , which is transmitted to humans through bites and scratches from infected cats and dogs. CSD occurs mainly in children and adolescents and is associated with different clinical signs [1–3]. Although the typical clinical feature is regional lymphadenopathy, atypical manifestations may also occur (i.e.…”

Section: Introductionmentioning

confidence: 99%

“…Although the typical clinical feature is regional lymphadenopathy, atypical manifestations may also occur (i.e. prolonged fever without lymphadenopathy), accompanied by systemic symptoms such as endocarditis, hepatic/splenic granuloma, neuroretinitis, and encephalopathy [1,4–8].…”

Section: Introductionmentioning

confidence: 99%

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Serological and molecular detection of Bartonella henselae in specimens from patients with suspected cat scratch disease in Italy: A comparative study

et al. 2019

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Cat scratch disease (CSD) is an infectious disease caused by Bartonella henselae, usually characterized by self-limiting regional lymphadenopathy and fever. Given the low clinical diagnostic sensitivity and specificity of conventional anti-B. henselae indirect immunofluorescence assays (IFAs), real-time polymerase chain reaction (PCR)-based detection of B. henselae is now being proposed as a more sensitive tool to diagnose CSD. Thus, here we have assessed the efficacy of real-time PCR in detecting B. henselae in different specimens from patients with suspected CSD and compared it to that of IFA. From March 2011 to May 2016, at the Microbiology and Virology Unit, Azienda Ospedaliera Universitaria Città della Salute e della Scienza di Torino, Turin, Italy, 115 clinical specimens (56 aspirated pus, 39 fresh lymph node biopsies, and 20 whole blood samples) and 99 sera from 115 patients with suspected CSD (62 females and 53 males between the ages of 3 months and 68 years) were analyzed by both real-time PCR, used in a qualitative way, and IFA (IgM and IgG) for the presence of B. henselae. For 16 patients, serological results were not available due to a clinical decision not to request the test. B. henselae DNA positivity was detected by real-time PCR in 37.39% of patients, while 62.61% of them were negative. Thus, patients were divided into two groups: real-time PCR+ (n = 43) and real-time PCR- (n = 72). Real-time PCR screening of whole blood, biopsies, and aspirated pus revealed B. henselae positivity in 40%, 38.46%, and 35.71% of patients, respectively. When we analyzed samples by IFA, we found the presence of B. henselae in 28 out of 99 (28.28%) patients, of which 11 (11.11%) belonged to the real-time PCR+ group and 17 (17.17%) to the real-time PCR- group. Among the 71 seronegative subjects, 16 (16.16%) were found positive for B. henselae by real-time PCR. Thus, by combining the results of both assays, we were able to increase the percentage of B. henselae positive specimens from 27.27% (real-time PCR) or 28.28% (IFA) to 44.44% (real-time PCR+IFA). Altogether, these findings indicate that the early detection of B. henselae in patients with suspicious CSD through combined real-time PCR and serological analyses can lead to a more accurate diagnosis of CSD, thereby allowing prompt and appropriate disease management.

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

Cited by 22 publications

References 18 publications

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

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

Utility of Molecular Identification and Quantitation of Bartonella Species with Species-Specific Real-Time PCR for Monitoring Treatment Response: A Case Series

Serological and molecular detection of Bartonella henselae in specimens from patients with suspected cat scratch disease in Italy: A comparative study

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