通过直接基因组测序发现的巨细胞病毒主要立即早期基因的变异
Variations in the cytomegalovirus major immediate-early gene found by direct genomic sequencing.
作者信息
Brytting M, Wahlberg J, Lundeberg J, Wahren B, Uhlén M, Sundqvist V A
机构信息
Department of Virology, Karolinska Institute, Stockholm, Sweden.
出版信息
J Clin Microbiol. 1992 Apr;30(4):955-60. doi: 10.1128/jcm.30.4.955-960.1992.
Abstract
An assay to detect and sequence DNA from human cytomegalovirus (HCMV) immediate-early gene region 1 has been developed; it involves in vitro amplification by the polymerase chain reaction and direct solid-phase sequencing of the amplified material. Urine samples from 16 patients tested positive for HCMV DNA in both a colorimetric assay for the detection of immobilized amplified nucleic acids and a standard polymerase chain reaction assay with agarose gel electrophoresis. Ten urine samples from healthy people tested negative in the same assays. Analysis of 106-bp fragments from seven patients and two laboratory HCMV strains (Ad 169 and Towne) demonstrated that the viral sequences were conserved in samples collected at different times from the same patient and in tissue-cultured samples. Two of the patient strains had variations in the amplified region, with a total of seven nucleotide substitutions yielding five amino acid alterations in the coding sequence.
摘要
已开发出一种用于检测人巨细胞病毒（HCMV）立即早期基因区域1的DNA并对其进行测序的检测方法；该方法包括通过聚合酶链反应进行体外扩增以及对扩增产物进行直接固相测序。在用于检测固定化扩增核酸的比色测定法和采用琼脂糖凝胶电泳的标准聚合酶链反应测定法中，16例患者的尿液样本检测出HCMV DNA呈阳性。10例健康人的尿液样本在相同检测中呈阴性。对7例患者以及两种实验室HCMV毒株（Ad 169和Towne）的106bp片段进行分析表明，在同一患者不同时间采集的样本以及组织培养样本中，病毒序列是保守的。其中两例患者毒株在扩增区域存在变异，共有七个核苷酸替换，导致编码序列中有五个氨基酸改变。
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