Scott-Taylor T H, Hammond G W
Cadham Provincial Laboratory, University of Manitoba, Winnipeg, Canada.
J Clin Microbiol. 1992 Jul;30(7):1703-10. doi: 10.1128/jcm.30.7.1703-1710.1992.
The application of DNA hybridization directly to clinical specimens has the potential of improving the diagnosis of fastidious types of adenovirus. In this study, the genome of one adenovirus type from each human subgenus (A to F) was systematically evaluated by hybridization for homologous sequences to find the optimal common probe for detection of all human adenovirus types. The area of cross-hybridization, most closely defined with adenovirus type 2 (Ad2), mapped from map units 11.4 to 16.1 and 26.9 to 29.7 and, principally, to a central area of the genome between map units 47.5 and 65.2. The last area, enclosing the hexon gene, was highly conserved. Cloned probes generated from this area demonstrated the greatest homology to heterologous types by hybridization analysis. A HindIII-BglII clone containing the hexon gene of Ad2 within narrow confines reacted most evenly with all adenoviral types and detected the DNA of all other subgenera with a sensitivity 2 logs greater than that of a complete genomic Ad2 probe. The most homologous adenoviral gene sequences were observed in genes involved with DNA replication or intimately connected to the hexon in the early capsid formation. These results show that the hexon gene constitutes the best single region of the adenovirus genome for use as a genus-specific probe for the diagnosis of all human adenoviral subgenera by DNA hybridization.
将DNA杂交直接应用于临床标本有改善对难培养型腺病毒诊断的潜力。在本研究中,通过杂交系统评估了来自每个人亚属(A至F)的一种腺病毒的基因组,以寻找用于检测所有人腺病毒类型的最佳通用探针。与2型腺病毒(Ad2)最紧密界定的交叉杂交区域,定位于图谱单位11.4至16.1以及26.9至29.7,主要位于图谱单位47.5至65.2之间的基因组中心区域。最后一个区域包含六邻体基因,高度保守。从该区域产生的克隆探针通过杂交分析显示与异源类型具有最大同源性。一个在狭窄范围内包含Ad2六邻体基因的HindIII - BglII克隆与所有腺病毒类型反应最均匀,并且检测所有其他亚属的DNA的灵敏度比完整基因组Ad2探针高2个对数。在参与DNA复制或在早期衣壳形成中与六邻体紧密相连的基因中观察到最同源的腺病毒基因序列。这些结果表明,六邻体基因构成腺病毒基因组中最佳的单一区域,可作为通过DNA杂交诊断所有人腺病毒亚属的属特异性探针。