新的定量微阵列抗体捕获（Q-MAC）测定鉴定了HBV感染的蒙古人

StephenW

Hepatology. 2016 Nov 23. doi: 10.1002/hep.28957. [Epub ahead of print]
A novel quantitative microarray antibody capture (Q-MAC) assay identifies an extremely high HDV prevalence amongst HBV infected Mongolians.Chen X1,2, Oidovsambuu O3,4, Liu P1, Grosely R1, Elazar M1, Winn VD5, Fram B1, Boa Z6, Dai H6, Dashtseren B3,7,8, Yagaanbuyant D3,7,8, Genden Z3,7, Dashdorj N7, Bungert A7, Dashdorj N3,7, Glenn JS1,9,10.
Author information

• 1Department of Medicine, Division of Gastroenterology and Hepatology, Stanford University School of Medicine, Palo Alto, California.
• 2Department of Infectious Disease, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
• 3Liver Center, Ulaanbaatar, Mongolia.
• 4Mongolian National University.
• 5Department of Obstetrics and Gynecology, Stanford University School of Medicine, Palo Alto, California.
• 6Department of Chemistry, Stanford University, Palo Alto, California.
• 7Onom Foundation, Ulaanbaatar, Mongolia.
• 8Mongolian National University of Health Sciences.
• 9Department of Microbiology and Immunology, Stanford University School of Medicine, Palo Alto, California.
• 10Veterans Administration Medical Center, Palo Alto, California.
  

AbstractHepatitis delta virus (HDV) causes the most severe form of human viral hepatitis. HDV requires a hepatitis B virus (HBV) co-infection to provide HDV with HBV surface antigen envelope proteins. The net effect of HDV is to make the underlying HBV disease worse, including higher rates of hepatocellular carcinoma (HCC). Accurate assessments of current HDV prevalence have been hampered by the lack of readily available and reliable quantitative assays, combined with the absence of an FDA-approved therapy. We sought to develop a convenient assay for accurately screening populations and to use this assay to determine HDV prevalence in a population with abnormally high rates of HCC. We developed a high throughput quantitative microarray antibody capture (Q-MAC) assay for anti-HDV IgG wherein recombinant HDV delta antigen is printed by microarray on slides coated with a noncontinuous, nanostructured plasmonic gold film, enabling quantitative fluorescent detection of anti-HDV antibody in small aliquots of patient serum. This assay was then used to screen all HBV-infected patients identified in a large randomly selected cohort designed to represent the Mongolian population. We identified two quantitative thresholds of captured antibody that were 100% predictive of the sample either being positive on standard western blot, or harboring HDV RNA detectable by qPCR, respectively. Subsequent screening of the HBV-positive cohort revealed that a remarkable 57% were RNA positive and an additional 4% were positive on western blot alone.
CONCLUSIONS: The Q-MAC assay's unique performance characteristics make it ideal for population screening. Its application to the Mongolian HBsAg+ population reveals an apparent ∼60% prevalence of HDV co-infection amongst these HBV-infected Mongolian subjects, which may help explain the extraordinarily high rate of HCC in Mongolia. This article is protected by copyright. All rights reserved.

© 2016 by the American Association for the Study of Liver Diseases.

KEYWORDS: HDV; anti-HDV; plasmonic gold; protein microarray

PMID:27880976DOI:10.1002/hep.28957

StephenW

肝脏病学。 2016 Nov 23. doi：10.1002 / hep.28957。 [打印前的电子版]
新的定量微阵列抗体捕获（Q-MAC）测定鉴定了HBV感染的蒙古人中极高的HDV流行率。
Chen X1,2，Oidovsambuu O3,4，Liu P1，Grosely R1，Elazar M1，Winn VD5，Fram B1，Boa Z6，Dai H6，Dashtseren B3,7,8，Yagaanbuyant D3,7,8，Genden Z3,7， Dashdorj N7，Bungert A7，Dashdorj N3,7，Glenn JS1,9,10。
作者信息

    1，斯坦福大学医学院胃肠病学与肝病学系，加利福尼亚州帕洛阿尔托。
    2.上海交通大学附属第六人民医院传染病教研室，上海。
    3Liver Center，乌兰巴托，蒙古。
    4蒙古国立大学。
    5斯坦福大学医学院，加利福尼亚州帕洛阿尔托的妇产科。
    6斯坦福大学化学系，帕洛阿尔托，加利福尼亚州。
    7Onom基金会，乌兰巴托，蒙古。
    8蒙古国立健康科学大学。
    9斯坦福大学医学院，帕洛阿尔托，加利福尼亚微生物学和免疫学室。
    10退伍军人管理医疗中心，帕洛阿尔托，加利福尼亚州。

抽象

丁型肝炎病毒（HDV）引起最严重形式的人类病毒性肝炎。 HDV需要乙型肝炎病毒（HBV）共感染以提供具有HBV表面抗原包膜蛋白的HDV。 HDV的净效应是使基础性HBV疾病更糟，包括更高的肝细胞癌（HCC）率。目前HDV流行率的准确评估由于缺乏现成的可靠的定量测定法，以及缺乏FDA批准的治疗而受到阻碍。我们试图开发一个方便的测定准确筛选人群，并使用此测定法确定HDV患病率在异常高的HCC率高的人口。我们开发了用于抗HDV IgG的高通量定量微阵列抗体捕获（Q-MAC）测定，其中通过微阵列在涂覆有非连续的纳米结构等离子体金膜的载玻片上印刷重组HDVδ抗原，使得能够定量荧光检测抗HDV抗体在小等分的患者血清中。然后将该测定用于筛选在设计为代表蒙古人群的大随机选择的队列中鉴定的所有HBV感染的患者。我们确定了捕获的抗体的两个定量阈值，100％预测样品在标准蛋白质印迹上是阳性的，或分别携带可通过qPCR检测的HDV RNA。随后筛选HBV阳性队列显示，显着的57％是RNA阳性，另外4％在单独的蛋白质印迹上是阳性的。
结论：

Q-MAC测定的独特性能特征使其成为群体筛选的理想选择。其应用于蒙古的HBsAg +群体揭示了在这些HBV感染的蒙古受试者中HDV共感染的明显〜60％的流行，这可能有助于解释蒙古的HCC的极高的比率。本文受版权保护。版权所有。

©2016美国肝病研究协会。
关键词：

HDV;抗HDV;等离子体激元蛋白质微阵列

PMID：
    27880976
DOI：
    10.1002 / hep.28957