Development of a Specific and Sensitive Enzyme-Linked Immunosorbent Assay as an In Vitro Diagnostic Tool for Detection of Bartonella henselae Antibodies in Human Serum

开发特异性和灵敏性的酶联免疫吸附试验作为检测人类血清中汉赛巴尔通体抗体的体外诊断工具

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作者:Markus Jost, Andreas Latz, Wibke Ballhorn, Volkhard A J Kempf

Abstract

Bartonella henselae causes cat scratch disease and several other clinical entities. Infections with B. henselae are frequently occurring; however, the infection is only rarely diagnosed, mainly due to a lack of knowledge in the medical community. Microscopic immunofluorescence assays (IFA) are widely used for the serodiagnosis of B. henselae infections but are laborious and time-consuming, and interpretation is subjective. An easy and reliable method for the serological diagnosis of B. henselae infections is needed to overcome the shortcomings of the current IFA. Here, we report the development of an ELISA detecting human anti-B. henselae antibodies from serum samples. By separating the water-insoluble fraction of B. henselae Houston-1 via ion-exchange chromatography, 16 subfractions were generated and tested for immunoreactivity via line blotting. One particular fraction (fraction 24) was selected and spotted on ELISA plates using an industrial production platform. By use of well-characterized human sera from the strictly quality-controlled serum library of the German National Consiliary Laboratory for Bartonella infections, the sensitivity of this ELISA was 100% for PCR-proven infections and 76% for clinically suspected infections at a specificity of 93%. This ELISA is therefore a reliable high-throughput method allowing the serodiagnosis of B. henselae infections.

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