通过液滴数字PCR建立爱泼斯坦-巴尔病毒相关胃癌的筛查方法
Establishment of a Screening Method for Epstein-Barr Virus-Associated Gastric Carcinoma by Droplet Digital PCR.
作者信息
Shuto Takuya, Nishikawa Jun, Shimokuri Kanami, Yanagi Ayaka, Takagi Tatsuya, Takagi Fumiya, Miura Osamu, Iida Michihisa, Nagano Hiroaki, Takemoto Yoshihiro, Harada Eijiro, Suehiro Yutaka, Yamasaki Takahiro, Okamoto Takeshi, Sakaida Isao
机构信息
Faculty of Laboratory Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan.
Hofu Institute of Gastroenterology, 14-33 Ekiminami-machi, Hofu, Yamaguchi 747-0801, Japan.
出版信息
Microorganisms. 2019 Nov 29;7(12):628. doi: 10.3390/microorganisms7120628.
BACKGROUND
Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is classified as one of the molecular subtypes of gastric cancer. We used droplet digital polymerase chain reaction (ddPCR) to enable highly sensitive and quantitative detection of EBV.
METHODS
EBV-DNA load was calculated based on the copy number of the BamH1-W fragment of EBV by ddPCR, and the cut-off value of EBV-DNA load was set. We conducted both ddPCR and EBER1 ISH to examine whether their results coincided in 158 gastric cancer specimens of unknown EBV status. We prepared 26 biopsy specimens and 49 serum samples including EBVaGC and assayed them by ddPCR.
RESULTS
The median values of EBV-DNA load for EBVaGC and EBV-negative control were 17.0 and 0.00308, respectively. A cut-off value of 0.032 was determined for which the sensitivity was 1. Among the 158 gastric cancer specimens, 14 lesions were judged as EBV-positive by the 0.032 cut-off value determined by ddPCR. The results of ddPCR and EBER1 ISH were in complete agreement. Even when using a biopsy specimen as a sample for ddPCR, the EBV-DNA load of all EBVaGCs was larger than the cut-off value.
CONCLUSIONS
We established a new method of diagnosing EBVaGC from tissue samples by ddPCR.
背景
爱泼斯坦-巴尔病毒相关胃癌（EBVaGC）被归类为胃癌的分子亚型之一。我们使用数字液滴聚合酶链反应（ddPCR）实现对EBV的高灵敏度和定量检测。
方法
通过ddPCR基于EBV的BamH1-W片段的拷贝数计算EBV-DNA载量，并设定EBV-DNA载量的临界值。我们对158例EBV状态未知的胃癌标本同时进行了ddPCR和EBER1原位杂交，以检查它们的结果是否一致。我们制备了包括EBVaGC在内的26份活检标本和49份血清样本，并通过ddPCR进行检测。
结果
EBVaGC和EBV阴性对照的EBV-DNA载量中位数分别为17.0和0.00308。确定临界值为0.032，其灵敏度为1。在158例胃癌标本中，根据ddPCR确定的0.032临界值，有14个病变被判定为EBV阳性。ddPCR和EBER1原位杂交的结果完全一致。即使使用活检标本作为ddPCR的样本，所有EBVaGC的EBV-DNA载量均大于临界值。
结论
我们建立了一种通过ddPCR从组织样本中诊断EBVaGC的新方法。
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