用干血点测定对乙肝病毒的免疫力

Chijdem Ismailova, E. Golkocheva-Markova, T. Tenev, S. Krumova
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引用次数: 0

摘要

背景:干血斑(DBS)被用来研究乙型肝炎病毒(HBV)感染在流行地区和高危人群中的流行情况。然而,通过ELISA检测DBS中HBV血清学标志物尚未完全优化。本研究的目的是评估DBS卡作为ELISA存储基质时抗- hbs的稀释水平。材料与方法:采用ELISA法检测乙型肝炎表面抗原(anti-HBs)抗体。检测了以下标本:20例患者与20例DBS配对的血清标本;20名接种hbv疫苗的卫生保健工作者的血清样本与20个干血清斑点(DSS)配对;以及四种不同稀释度的免疫球蛋白。为了研究样品稀释问题,采用了不同的洗脱方案。结果:与“金标准”相比,DBS的特异性为100%,灵敏度为45%。对洗脱的DBS/DSS样品进行稀释,在某些情况下测得的抗hbs滴度降至10 mIU/ml以下。初始抗hbs血清滴度阳性与DBS/DSS检测结果无相关性。此外,在检测免疫球蛋白时,对洗脱的DSS样品测量20至50倍的稀释度。洗脱样品浓度增加,DSS抗hbs滴度升高。结论:由于DBS中少量样品滴度较低,为了解决稀释问题,有必要对不同的洗脱方案进行验证。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
USE OF DRIED BLOOD SPOTS FOR MEASURING THE IMMUNITY AGAINST HBV
Background: Dried blood spots (DBS) have been used to study the prevalence of hepatitis B virus (HBV) infection in endemic areas and in high-risk groups. However, detection of HBV serological markers in DBS by ELISA assays has not yet been fully optimised. The aim of the present study is to evaluate the dilution level of anti-HBs when DBS cards are used as storage matrix implemented for ELISA. Material and methods: Antibodies against hepatitis B surface antigen (anti-HBs) were detected by ELISA. The following specimens were examined: serum samples from 20 patients paired with 20 DBS; serum samples from 20 HBV-vaccinated healthcare workers paired with 20 dried serum spots (DSS); and four different dilutions of Immunovenin. Different elution protocols were used in order to study the problem with sample dilution. Results: Specificity of 100% and sensitivity of 45% were established for DBS versus the “gold standard”. Dilution of the eluted DBS/DSS samples was established and in some cases the measured anti-HBs titre dropped under 10 mIU/ml. Correlation was not observed between the positive initial anti-HBs serum titres and the obtained values of DBS/DSS testing. Also, 20- to 50-fold dilutions were measured for eluted DSS samples when testing Immunovenin. Increasing of the eluted sample concentration raised DSS anti-HBs titre. Conclusions: In order to resolve the problem of dilution, it is necessary to validate different elution protocols because the small amount of sample in DBS showed lower titres.
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