Fujii Hikaru, Kakiuchi Satsuki, Tsuji Masanori, Nishimura Hidekazu, Yoshikawa Tomoki, Yamada Souichi, Omura Natsumi, Inagaki Takuya, Shibamura Miho, Harada Shizuko, Taniguchi Shuichi, Saijo Masayuki
Department of Virology I, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
Department of Pediatrics, The University of Tokyo Hospital, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
J Virol Methods. 2018 Jan;251:123-128. doi: 10.1016/j.jviromet.2017.10.019. Epub 2017 Oct 23.
Ion Torrent next-generation sequencing (NGS) technology was applied to study the mode of emergence of acyclovir (ACV)-resistant (ACVr) herpes simplex virus type 1 (HSV-1) in patients with hematopoietic stem cell transplantation (HSCT) by quantitatively detecting mutations in the viral thymidine kinase (vTK) gene in the HSV-1 isolates recovered from HSCT patients. All of the mutations detected with the Sanger sequencing method in the vTK genes of HSV-1 isolates were also detected with the NGS assay. Furthermore, different mutations, which conferred ACV resistance and were not detected with the Sanger sequencing method, were also detected in a quantitative manner by using the NGS assay. The approach described here is applicable to studying the emergence process of vTK gene mutation-associated ACVr HSV-1 more in detail than the Sanger method. The NGS assay makes it possible to make a diagnosis of vTK gene mutation-associated ACVr HSV-1 infections at the early stage, which the ratio of ACVr HSV-1 is much lower than that of ACV-sensitive (ACVs) HSV-1.
采用离子激流下一代测序（NGS）技术，通过定量检测从造血干细胞移植（HSCT）患者中分离出的单纯疱疹病毒1型（HSV-1）毒株中病毒胸苷激酶（vTK）基因的突变，来研究HSCT患者中阿昔洛韦（ACV）耐药（ACVr）HSV-1的出现模式。用Sanger测序法在HSV-1分离株vTK基因中检测到的所有突变，也能用NGS检测法检测到。此外，通过使用NGS检测法，还能定量检测到一些用Sanger测序法未检测到的、赋予ACV耐药性的不同突变。与Sanger法相比，本文所述方法更适用于更详细地研究vTK基因突变相关的ACVr HSV-1的出现过程。NGS检测法能够在ACVr HSV-1比例远低于阿昔洛韦敏感（ACVs）HSV-1的早期阶段，诊断vTK基因突变相关的ACVr HSV-1感染。
