Scott-Taylor T H, Ahluwalia G, Hammond G W
Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada.
J Virol Methods. 1992 Jul;38(1):25-37. doi: 10.1016/0166-0934(92)90166-b.
Enteric adenoviruses 40 and 41 (Ad40 and Ad41) are a prominent cause of gastroenteritis in young children. Diagnosis of these enteric types by conventional means is complicated by their fastidious growth characteristics. Enteric adenovirus growth was enhanced by cocultivation. Typing of enteric isolates currently entails analysis of the extracted viral DNA with restriction enzymes. Restriction endonuclease fragments of the Ad41 strain Tak genome were ordered by (i) double digestion, (ii) release of restriction fragments from plasmids containing 84% of the Ad41 genome in EcoRI fragments A, B and C, (iii) hybridization of Southern blotted Ad41 fragments with EcoRI fragment containing plasmids and various segments of the Ad2 genome, (iv) sequential reduction of the genome beginning with terminal restriction fragments with exonuclease III and S1 nuclease. The termini of adenovirus genomes are difficult to clone and the use of exonuclease III is a useful alternative to conventional restriction mapping. DNA restriction patterns, fragment sizes and restriction maps of the Ad4 1 strain Tak with enzymes BamHI, BglII, ClaI, EcoRI, HindIII, PstI, SalI, SmaI and XhoI are presented. Prototype strain restriction maps should enable better understanding of adenovirus type 41 and its epidemiology.
肠道腺病毒40型和41型（Ad40和Ad41）是幼儿肠胃炎的主要病因。由于它们苛求的生长特性，通过传统方法诊断这些肠道型别很复杂。共培养可增强肠道腺病毒的生长。目前对肠道分离株进行分型需要用限制性酶分析提取的病毒DNA。Ad41 Tak株基因组的限制性内切酶片段通过以下方法排序：（i）双酶切；（ii）从含有Ad41基因组84%的EcoRI片段A、B和C的质粒中释放限制性片段；（iii）将Southern印迹的Ad41片段与含有EcoRI片段的质粒和Ad2基因组的各个片段杂交；（iv）从末端限制性片段开始用核酸外切酶III和S1核酸酶依次减少基因组。腺病毒基因组的末端难以克隆，使用核酸外切酶III是传统限制性图谱分析的一种有用替代方法。给出了Ad41 Tak株用BamHI、BglII、ClaI、EcoRI、HindIII、PstI、SalI、SmaI和XhoI酶切后的DNA限制性图谱、片段大小和限制性图谱。原型株限制性图谱应有助于更好地了解腺病毒41型及其流行病学。
