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. Conclusion: 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)

中文翻译：

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一种新颖的定量微阵列抗体捕获测定法可确定感染乙型肝炎病毒的蒙古人中极高的丙型肝炎三角洲病毒流行率

三角洲肝炎病毒（HDV）导致人类病毒性肝炎的最严重形式。HDV需要乙型肝炎病毒（HBV）共同感染才能为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，包括更高的肝细胞癌发病率。由于缺乏现成的，可靠的定量分析方法，以及缺乏美国食品药品监督管理局批准的治疗方法，目前对HDV流行的准确评估受到了阻碍。我们试图开发一种方便的测定方法，以准确筛查人群，并使用该测定方法确定肝细胞癌异常高发生率人群中的HDV患病率。我们针对抗HDV免疫球蛋白G开发了高通量定量微阵列抗体捕获测定法，其中重组HDVδ抗原通过微阵列印刷在涂有不连续，纳米结构的等离激元金膜的载玻片上，从而能够以小等份定量荧光检测抗HDV抗体患者血清。然后，该分析方法用于筛选在代表蒙古族人群的大型随机选择队列中鉴定出的所有HBV感染患者。我们确定了两个捕获到的抗体的定量阈值，这些阈值可以100％预测样品在标准Western印迹上呈阳性或包含可通过实时定量PCR检测到的HDV RNA。乙肝病毒的后续筛查 可以定量分析患者血清中的小份抗HDV抗体。然后，该分析方法用于筛选在代表蒙古族人群的大型随机选择队列中鉴定出的所有HBV感染患者。我们确定了两个捕获到的抗体的定量阈值，这些阈值可以100％预测样品在标准Western印迹上呈阳性或包含可通过实时定量PCR检测到的HDV RNA。乙肝病毒的后续筛查 可以定量分析患者血清中的小份抗HDV抗体。然后，该分析方法用于筛选在代表蒙古族人群的大型随机选择队列中鉴定出的所有HBV感染患者。我们确定了两个捕获到的抗体的定量阈值，这些阈值可以100％预测样品在标准Western印迹上呈阳性或包含可通过实时定量PCR检测到的HDV RNA。乙肝病毒的后续筛查 我们确定了两个捕获到的抗体的定量阈值，这些阈值可以100％预测样品在标准Western印迹上呈阳性或包含可通过实时定量PCR检测到的HDV RNA。乙肝病毒的后续筛查 我们确定了两个捕获到的抗体的定量阈值，这些阈值可以100％预测样品在标准Western印迹上呈阳性或包含可通过实时定量PCR检测到的HDV RNA。乙肝病毒的后续筛查⁺队列研究表明，单独的蛋白质印迹法显示，显着的57％是RNA ⁺，另外4％是阳性。结论：定量微阵列抗体捕获测定法的独特性能特征使其非常适合人群筛查。在蒙古HBV表面抗原阳性人群中的应用表明，在这些被HBV感染的蒙古族受试者中，HDV合并感染的患病率约为60％，这可能有助于解释蒙古肝细胞癌的异常高发率。（肝病学， 2017年； 66：1739–1749）