Huang C, Deibel R
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
J Clin Microbiol. 1988 Dec;26(12):2652-6. doi: 10.1128/jcm.26.12.2652-2656.1988.
A number of recombinant plasmids containing genomic segments of adenovirus were constructed. Seven cloned probes, as well as total adenovirus type 2 (Ad2) and Ad16 genomic DNA, were tested by a nucleic acid hybridization technique for sensitivity and specificity in detecting adenoviruses in infected cells. Adenovirus DNA was spotted onto a nitrocellulose filter and hybridized with 32P-labeled DNA probes. The probes, total Ad2 genomic DNA, and plasmid pAd2-H (containing the hexon gene from Ad2 DNA) all detected 10 reference serotypes of five genomic subgroups (A through E) with similar sensitivities. However, plasmid pAd2-H required less preparation time than did total Ad2 DNA. Probes pAd2-F (containing the fiber gene from Ad2) and pAd16-BD (containing the BamHI D fragment from Ad16) hybridized only with reference serotypes from the homologous subgroups (C and B, respectively). Of 101 patient isolates amplified in cells, pAd2-H detected 100% of all isolates from both the homologous and the heterologous subgroups. The detection rates for pAd2-F were 100% (subgroup C) and 3.6% (subgroups A, B, and D), and those for pAd16-BD were 100% (subgroup B) and 9.4% (subgroups A, C, and D). A commercial biotinylated product (Pathogene II) was also included in this study for comparison.
构建了许多含有腺病毒基因组片段的重组质粒。通过核酸杂交技术对7种克隆探针以及2型腺病毒（Ad2）和16型腺病毒（Ad16）的基因组DNA进行检测，以评估其在检测感染细胞中腺病毒时的敏感性和特异性。将腺病毒DNA点样到硝酸纤维素滤膜上，与用³²P标记的DNA探针杂交。这些探针、Ad2基因组总DNA以及质粒pAd2-H（含有来自Ad2 DNA的六邻体基因）均能以相似的敏感性检测五个基因组亚群（A至E）的10种参考血清型。然而，与Ad2总DNA相比，质粒pAd2-H所需的制备时间更短。探针pAd2-F（含有来自Ad2的纤维基因）和pAd16-BD（含有来自Ad16的BamHI D片段）仅分别与同源亚群（分别为C和B）的参考血清型杂交。在细胞中扩增的101株患者分离株中，pAd2-H检测到同源和异源亚群的所有分离株的比例均为100%。pAd2-F对C亚群的检测率为100%，对A、B和D亚群的检测率为3.6%；pAd16-BD对B亚群的检测率为100%，对A、C和D亚群的检测率为9.4%。本研究还纳入了一种商业生物素化产品（Pathogene II）进行比较。
