Analysis of ganciclovir resistant HHV-6B clinical isolates by using quenching probes PCR (QP-PCR) methodology.
Analysis of ganciclovir resistant HHV-6B clinical isolates by using quenching probes PCR (QP-PCR) methodology.
复制标题
使用猝灭探针 PCR (QP-PCR) 方法分析更昔洛韦耐药 HHV-6B 临床分离株。
DOI：
10.1128/aac.04692-14
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发表时间：
2015
影响因子：
4.9
通讯作者：
Yoshikawa T
中科院分区：
文献类型：
--
作者：
Hiramatsu H;Suzuki R;Yamada S;Ihira M;Kawamura Y;Matsuoka E;Miura H;Isegawa Y;Yoshikawa T
采用猝灭探针PCR （QP-PCR）方法测定从原发病毒感染和病毒再激活患者中分离的人疱疹病毒6B （HHV-6B）临床分离株更昔洛韦（GCV）耐药频率。从15例造血干细胞移植（HSCT）患者中反复分离出42株HHV-6B临床分离株，从20例exanthem subitum （ES）患者中分离出20株。在15例HSCT受者中，9例在观察期间接受了GCV；然而，没有ES患者接受GCV治疗。以已建立的实验室菌株Z29和HST为标准菌株。所有临床分离株的U69基因1区和2区与Z29株的1区和2区熔解温度相同。在第3区，所有临床分离株的熔点温度介于含有A462D单核苷酸多态性（SNP）的质粒的熔点温度和Z29菌株的熔点温度之间，所有临床分离株的熔点温度分布与日本HST菌株的熔点温度分布相似。正如预期的那样，从ES患者中恢复的20株临床分离株和从未接受GCV治疗的HSCT受体中恢复的14株分离株均未携带与GCV耐药相关的6个已知snp。有趣的是，这6个snp在9例接受GCV移植的HSCT患者的28个临床分离株中未被检测到。对来自15例HSCT受者的15个代表性分离株的U69基因进行额外的序列分析，发现了其他snp。这些snp与在HST菌株中发现的相同。因此，抗GCV的HHV-6B毒株的出现率似乎相对较低，即使在接受GCV治疗的HSCT受体中也是如此。
Quenching probe PCR (QP-PCR) analysis was used to determine the frequency of ganciclovir (GCV) resistance among clinical isolates of human herpesvirus 6B (HHV-6B) obtained from patients with primary viral infection and viral reactivation. Forty-two HHV-6B clinical isolates were repeatedly recovered from 15 hematopoietic stem cell transplant (HSCT) recipients, and 20 isolates were recovered from 20 exanthem subitum (ES) patients. Of the 15 HSCT recipients, 9 received GCV during the observation period; however, none of the ES patients were treated with GCV. Two established laboratory strains, Z29 and HST, were used as standards in this study. Regions 1 and 2 of the U69 gene of all of the clinical isolates demonstrated the same melting temperature as regions 1 and 2 of the Z29 strain. For region 3, the melting temperatures of all clinical isolates fell between the melting temperature of the plasmid containing the A462D single nucleotide polymorphism (SNP) and the melting temperature of the Z29 strain, and the melting temperatures profiles of all clinical isolates were similar to the melting temperature profile of the Japanese HST strain. As expected, none of the 20 clinical isolates recovered from the ES patients and the 14 isolates recovered from the HSCT recipients who did not receive GCV treatment carried the six known SNPs associated with GCV resistance. Interestingly, these six SNPs were not detected in the 28 clinical isolates recovered from the 9 HSCT recipients who received GCV. Additional sequence analysis of the U69 gene from the 15 representative isolates from the 15 HSCT recipients identified other SNPs. These SNPs were identical to those identified in the HST strain. Therefore, the rate of emergence of GCV-resistant HHV-6B strains appears to be relatively low, even in HSCT recipients treated with GCV.