Chen Xiaohua, Oidovsambuu Odgerel, Liu Ping, Grosely Rosslyn, Elazar Menashe, Winn Virginia D, Fram Benjamin, Boa Zhang, Dai Hongjie, Dashtseren Bekhbold, Yagaanbuyant Dahgwahdorj, Genden Zulkhuu, Dashdorj Naranbaatar, Bungert Andreas, Dashdorj Naranjargal, Glenn Jeffrey S
Department of Medicine, Division of Gastroenterology and Hepatology, Stanford University School of Medicine, Palo Alto, CA.
Liver Center, Ulaanbaatar, Mongolia.
Hepatology. 2017 Dec;66(6):1739-1749. doi: 10.1002/hep.28957. Epub 2017 Oct 30.
Hepatitis delta virus (HDV) causes the most severe form of human viral hepatitis. HDV requires a hepatitis B virus (HBV) coinfection 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. Accurate assessments of current HDV prevalence have been hampered by the lack of readily available and reliable quantitative assays, combined with the absence of a Food and Drug Administration-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 hepatocellular carcinoma. We developed a high-throughput quantitative microarray antibody capture assay for anti-HDV immunoglobulin G 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 real-time quantitative PCR. Subsequent screening of the HBV cohort revealed that a remarkable 57% were RNA and an additional 4% were positive on western blot alone.
The quantitative microarray antibody capture assay's unique performance characteristics make it ideal for population screening; its application to the Mongolian HBV surface antigen-positive population reveals an apparent ∼60% prevalence of HDV coinfection among these HBV-infected Mongolian subjects, which may help explain the extraordinarily high rate of hepatocellular carcinoma in Mongolia. (Hepatology 2017;66:1739-1749).
丁型肝炎病毒（HDV）导致人类最严重的病毒性肝炎形式。HDV需要与乙型肝炎病毒（HBV）合并感染，以便为HDV提供HBV表面抗原包膜蛋白。HDV的净效应是使潜在的HBV疾病恶化，包括肝细胞癌的更高发病率。由于缺乏现成且可靠的定量检测方法，再加上没有食品药品监督管理局批准的治疗方法，目前对HDV流行率的准确评估受到了阻碍。我们试图开发一种方便的检测方法来准确筛查人群，并使用该检测方法来确定在肝细胞癌发病率异常高的人群中HDV的流行率。我们开发了一种用于抗HDV免疫球蛋白G的高通量定量微阵列抗体捕获检测方法，其中重组HDVδ抗原通过微阵列印在涂有不连续纳米结构等离子体金膜的载玻片上，能够对患者血清小样本中的抗HDV抗体进行定量荧光检测。然后使用该检测方法对在一个旨在代表蒙古人群体的大型随机选择队列中识别出的所有HBV感染患者进行筛查。我们确定了捕获抗体的两个定量阈值，这两个阈值对样本在标准蛋白质印迹上呈阳性或含有可通过实时定量PCR检测到的HDV RNA具有100%的预测性。随后对HBV队列的筛查显示，显著的57%为RNA阳性，另外4%仅在蛋白质印迹上呈阳性。
定量微阵列抗体捕获检测方法独特的性能特征使其非常适合人群筛查；将其应用于蒙古HBV表面抗原阳性人群，揭示了在这些HBV感染的蒙古受试者中HDV合并感染的明显患病率约为60%，这可能有助于解释蒙古肝细胞癌的极高发病率。（《肝脏病学》2017年；66:1739 - 1749）
