Ohye Tamae, Kawamura Yoshiki, Inagaki Hidehito, Yoshikawa Akiko, Ihira Masaru, Yoshikawa Tetsushi, Kurahashi Hiroki
Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan; Department of Clinical Hematology, Faculty of Medical Technology, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
J Virol Methods. 2016 Feb;228:74-8. doi: 10.1016/j.jviromet.2015.11.001. Epub 2015 Nov 6.
Some healthy individuals carry human herpesvirus-6 (HHV-6) within a host chromosome, which is called inherited chromosomally integrated human herpesvirus-6 (iciHHV-6). Because iciHHV-6 is generally considered a non-pathogenic condition, it is important to distinguish iciHHV-6 from HHV-6 reactivation in immunocompromised hosts because both conditions manifest high copy numbers of the HHV-6 in peripheral blood mononuclear cells. Although fluorescent in situ hybridization (FISH) is a reliable method for the diagnosis of iciHHV-6, HHV-6-specific FISH probes are not commercially available. In our present study, we established a simple PCR-based method for producing FISH probes that can detect the chromosomal integration site of iciHHV-6 at high sensitivity. Using these probes, we confirmed that HHV-6 signals were consistently located at the telomeric region in all of the 13 iciHHV-6 individuals examined. Interestingly, in all seven Japanese iciHHV-6A patients, signals were detected exclusively on chromosome 22q. This method provides a simple and fast approach for iciHHV-6 diagnosis in the clinical laboratory.
一些健康个体在宿主染色体中携带人类疱疹病毒6型（HHV-6），这被称为遗传性染色体整合人类疱疹病毒6型（iciHHV-6）。由于iciHHV-6通常被认为是一种非致病性状态，因此区分iciHHV-6与免疫功能低下宿主中的HHV-6再激活很重要，因为这两种情况在外周血单个核细胞中均表现为HHV-6的高拷贝数。尽管荧光原位杂交（FISH）是诊断iciHHV-6的可靠方法，但HHV-6特异性FISH探针尚无商业供应。在我们目前的研究中，我们建立了一种基于PCR的简单方法来生产FISH探针，该探针能够高灵敏度地检测iciHHV-6的染色体整合位点。使用这些探针，我们证实，在所检测的13例iciHHV-6个体中，HHV-6信号始终位于端粒区域。有趣的是，在所有7例日本iciHHV-6A患者中，信号仅在22号染色体长臂上被检测到。该方法为临床实验室中iciHHV-6的诊断提供了一种简单快速的途径。
