Rapid and sensitive identification of glycoprotein H genotypes in clinical human cytomegalovirus samples.
Rapid and sensitive identification of glycoprotein H genotypes in clinical human cytomegalovirus samples.
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DOI：
10.7883/yoken.jjid.2014.017
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发表时间：
2015
影响因子：
2.2
通讯作者：
Wei Li;R. Tao;Xiao Zhang;Q. Shu;Hui-hui Gao;S. Shang;Zhao-Yang Peng;Huamei Li
中科院分区：
文献类型：
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作者：
Wei Li;R. Tao;Xiao Zhang;Q. Shu;Hui-hui Gao;S. Shang;Zhao-Yang Peng;Huamei Li
人巨细胞病毒（HCMV）是一种常见的病原体，可引起免疫缺陷患者的持续感染，并导致显著的发病率和死亡率，特别是在移植受者和儿童中。不同的HCMV糖蛋白H（gH）基因型可能导致不同的疾病并影响这些疾病的严重程度。为了建立一种灵敏的实时荧光定量PCR检测方法，可以快速区分两种HCMV gH基因型，引物设计的目标是gH基因的保守区域。设计gH 1和gH 2探针以靶向两个可变区。本研究使用HCMV标准株（AD 169和TOWNE）和203例HCMV感染儿童的临床尿液样本。基于用于检测HCMV的靶gH基因片段的引物-探针组，我们的定量实时PCR检测特异性区分HCMV gH 1和gH 2，检测限约为10（2）病毒拷贝/ml。在检测的203份临床尿液样本中，145份gH 1阳性，56份gH 2阳性，2份两者均阳性。因此，我们开发了一种基于gH基因的真实的时间PCR方法，可以快速，稳定，特异性区分两种HCMV gH基因型。我们发现HCMV gH 1是常见的儿童检查在浙江，中国。
Human cytomegalovirus (HCMV) is a common pathogen that causes persistent infections in immune deficient patients and results in significant morbidity and mortality, particularly among transplant recipients and children. Different HCMV glycoprotein H (gH) genotypes may cause different diseases and affect the severity of these diseases. To develop a sensitive quantitative real-time PCR assay that could rapidly distinguish between two HCMV gH genotypes, primers were designed to target the conserved region of the gH gene. gH1 and gH2 probes were designed to target the two variable regions. Standard HCMV strains (AD169 and TOWNE) and 203 clinical urine samples from HCMV infected children were used for the present study. Based on the primer-probe set used to detect the target gH gene segment of HCMV, our quantitative real-time PCR assay specifically discriminated between HCMV gH1 and gH2 with a detection limit of approximately 10(2) viral copies/ml. Among the 203 clinical urine samples tested, 145 were gH1 positive, 56 were gH2 positive, and 2 were positive for both. Thus, we developed a gH gene-based real time-PCR method that could rapidly, stably, and specifically distinguish between two HCMV gH genotypes. We found HCMV gH1 to be common among children examined in Zhejiang, China.