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Enriched high-titer anti-hepatitis A virus (HAV) IgM for positive controls in diagnostic devices.

机译：丰富的高滴度抗A型肝炎病毒（HAV）IgM，可用于诊断设备中的阳性对照。

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摘要

Hepatitis A refers to liver inflammation caused by infection with the hepatitis A virus (HAV). The HAV is one of several viruses that can cause hepatitis and is one of the 3 most common hepatitis viruses in the United States. The other 2 are hepatitis B and hepatitis C viruses. The presence of anti-HAV IgM in serum indicates the acute or early convalescent stage of HAV infection. Although several commercial tests are available to aid in diagnosis, the lack of available high titer plasma for positive control, due to the short window of the IgM response following infection (5--10 weeks after exposure) and the low U.S. incidence of the HAV (one in 20,000 or 5 cases per 100,000) is a persistent problem. Samples collected prior to 4 and later than 12 weeks from exposure to HAV often demonstrate anti-HAV IgM titers too low to be useful as a positive control. A low titer stock of anti-hepatitis A (HAV-IgM) with a signal-to-cut-off ratio of &sim3 is abundantly available. It requires at least a 5-fold enrichment before it can be used in medical diagnostic devices. In order to achieve this goal, we have developed a simple two-step process for enrichment. The first step consists of a fractionation of plasma proteins by ammonium sulfate (A/S) precipitation. The second step is the use of a Pellicon concentrator of molecular weight cut-off of 500 kD. Identity of the enriched anti-HAV IgM product was evaluated by SDS-PAGE and Western blotting. The functionality of IgM was evaluated by ELISA, Abbott AxSYM HAVAB-M 2.0, Abbott AxSYM HAVAB 2.0, DiaSorin ETI-HA-IGMK PLUS, and DiaSorin ETI-AB-HAVK PLUS assays. The enriched anti-HAV IgM was formulated in normal human plasma and was found to be stable for 11 days at 37°C. This was translated to 15 months of stability at 2--8°C by the Arrhenius plotting model (Kannen 1964). In addition to the enriched anti-HAV IgM product, the process should be amenable to production of IgM-positive controls from low-titer anti-HBcore IgM, anti-Toxoplasma IgM, and anti-CMV IgM plasma, as well as from other disease-state plasma where high titer IgM plasma is in short supply.
机译：甲型肝炎是指由甲型肝炎病毒（HAV）感染引起的肝脏炎症。 HAV是可能引起肝炎的几种病毒之一，并且是美国3种最常见的肝炎病毒之一。其他2种是乙型肝炎和丙型肝炎病毒。血清中存在抗HAV IgM，表明HAV感染处于急性或恢复期。尽管有几种商业测试可用于帮助诊断，但由于感染后（暴露后5--10周）IgM反应时间短且美国HAV发生率较低，因此缺乏可用于阳性对照的高滴度血浆（20,000例中的一个或每100,000中5例）是一个持续存在的问题。暴露于HAV之前4周和之后12周之后收集的样品通常显示出抗HAV IgM滴度太低，无法用作阳性对照。低滴度的抗甲型肝炎病毒（HAV-IgM）的信噪比为＆sim3。它至少需要浓缩5倍，才能用于医疗诊断设备。为了实现此目标，我们开发了一个简单的两步富集过程。第一步包括通过硫酸铵（A / S）沉淀分离血浆蛋白。第二步是使用截留分子量为500 kD的Pellicon浓缩器。通过SDS-PAGE和蛋白质印迹评估富集的抗HAV IgM产物的身份。通过ELISA，Abbott AxSYM HAVAB-M 2.0，Abbott AxSYM HAVAB 2.0，DiaSorin ETI-HA-IGMK PLUS和DiaSorin ETI-AB-HAVK PLUS分析评估了IgM的功能。在正常人血浆中配制了丰富的抗HAV IgM，发现在37℃下稳定11天。通过Arrhenius绘图模型（Kannen 1964）将其转换为在2--8°C下15个月的稳定性。除了丰富的抗HAV IgM产品外，该过程还应适用于从低滴度抗HBcore IgM，抗弓形虫IgM和抗CMV IgM血浆以及其他疾病中产生IgM阳性对照状态血浆，其中高滴度IgM血浆供不应求。

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  • 作者

    Patamawenu, Terry.;

  • 作者单位

    Hood College.;

  • 授予单位 Hood College.;
  • 学科 Biology Molecular.Health Sciences Immunology.Engineering Biomedical.
  • 学位 M.S.
  • 年度 2010
  • 页码 99 p.
  • 总页数 99
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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