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微小巴贝斯虫特异性免疫显性表位的鉴定及用于检测血清中抗体的肽酶免疫测定法的开发。

Identification of Babesia microti-specific immunodominant epitopes and development of a peptide EIA for detection of antibodies in serum.

作者信息

Houghton Raymond L, Homer Mary J, Reynolds Lisa D, Sleath Paul R, Lodes Michael J, Berardi Victor, Leiby David A, Persing David H

机构信息

Department of Antigen Discovery, Corixa Corporation, Seattle, Washington 98104, USA.

出版信息

Transfusion. 2002 Nov;42(11):1488-96. doi: 10.1046/j.1537-2995.2002.00215.x.

Abstract

BACKGROUND

Babesia microti is a tick-borne agent that is increasingly implicated in transfusion-acquired infection, especially in immunocompromised and elderly recipients. To develop a test that can detect antibody responses to B. microti, peptide epitopes identified in two serocomplementary B. microti-specific antigens were used in a prototype EIA.

STUDY DESIGN AND METHODS

A prototype peptide EIA was used to detect B. microti-specific antibodies in 15 sera taken before infection and 107 taken after infection from 59 individuals with known tick-borne infections previously confirmed by other methods. Three additional groups of samples were also tested: a proficiency panel of 18 sera positive for B. microti by IFA, 38 sera from blood donors confirmed positive by IFA, and 30 sera from random blood donors.

RESULTS

The combination peptide detected 98 out of 107 sera taken after infection that were IgG blot positive (4 equivocal). This included all 12 samples that were PCR positive and six sera from smear-negative patients that were confirmed positive by PCR, immunoblot, or IFA. Of the IgG blot-positive specimens that were equivocal (four specimens) or did not react (nine specimens) by EIA, most had low IFA titers consistent with previous exposure. In a second evaluation, 15 out of 15 Babesia IFA-positive sera and 3 out of 3 Babesia-Ehrlichia IFA-positive sera were positive, whereas sera from 30 random donors were negative. Finally, of 38 IFA-positive blood-donor samples, 35 were positive by peptide EIA. The three EIA-negative sera were Western blot negative.

CONCLUSION

Reactivity of the B. microti-specific peptide EIA shows a high correlation with IFA, PCR, and B. microti immunoblot in confirmed B. microti cases. The peptide EIA may be the most suitable B. microti infection test for adaptation to the blood bank environment if testing for B. microti is required in the future.

摘要

背景

微小巴贝斯虫是一种蜱传播病原体,越来越多地与输血获得性感染有关,尤其是在免疫功能低下者和老年受血者中。为开发一种能够检测针对微小巴贝斯虫抗体反应的检测方法,在一种原型酶免疫测定法中使用了在两种血清互补的微小巴贝斯虫特异性抗原中鉴定出的肽表位。

研究设计与方法

使用一种原型肽酶免疫测定法检测来自59名先前经其他方法确诊为蜱传播感染的个体的15份感染前血清和107份感染后血清中的微小巴贝斯虫特异性抗体。还检测了另外三组样本:一组由18份间接荧光抗体试验(IFA)检测微小巴贝斯虫呈阳性的血清组成的能力验证样本组、38份经IFA确认呈阳性的献血者血清以及30份随机献血者血清。

结果

组合肽在107份感染后IgG印迹呈阳性(4份结果不明确)的血清中检测出98份。这包括所有12份PCR呈阳性的样本以及6份涂片阴性患者的血清,这些血清经PCR、免疫印迹或IFA确认为阳性。在酶免疫测定法中结果不明确(4份样本)或无反应(9份样本)的IgG印迹阳性标本中,大多数IFA滴度较低,与既往接触情况相符。在第二次评估中,15份巴贝斯虫IFA阳性血清中的15份以及3份巴贝斯虫 - 埃立克体IFA阳性血清中的3份呈阳性,而30份随机献血者的血清呈阴性。最后,在38份IFA阳性的献血者样本中,35份经肽酶免疫测定法检测呈阳性。3份酶免疫测定法阴性的血清Western印迹呈阴性。

结论

在确诊的微小巴贝斯虫病例中,微小巴贝斯虫特异性肽酶免疫测定法的反应性与IFA、PCR以及微小巴贝斯虫免疫印迹高度相关。如果未来需要检测微小巴贝斯虫,肽酶免疫测定法可能是最适合应用于血库环境的微小巴贝斯虫感染检测方法。

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