Uryvaev L V, Rusavskaia E A, Sinagatullina N M, Faĭzullin L Z, Vengerov Iu Iu, Parasiuk N A, Ionova K S, Veĭko N N, Spitkovskiĭ D M, Karpukhin A V
Vopr Virusol. 1990 Nov-Dec;35(6):464-6.
A probe containing full-size DNA copy of influenza A/USSR/90/70 virus protein gene M labeled with biotin on 32P was used for influenza A virus RNA detection by dot hybridization method. For labeling with biotin, a new method of its administration by chemical modification of nucleic acid was employed. In homologous DNA:DNA hybridization the sensitivity of determinations was less than 1 pg in the biotin-treatment of the probe and 1.25 pg in its radioactive labeling. Hybridization of DNA probe with cytoplasmic RNA isolated from influenza A virus-infected (strains A/USSR/90/77 and A/Texas/77) MDCK cells revealed RNA in the dot corresponding to 4.5-5.5 1g ID50 of virus present in 2 x 10(4) cells. The probe did not bind with negative controls in any dot in all the tests. The results of the study indicate that DNA probes labeled with biotin and 32P and used in dot hybridization for influenza A virus RNA detection in infected cells show the similar sensitivity and specificity.
用生物素标记的含有甲型流感病毒A/苏联/90/70株病毒蛋白基因M的全尺寸DNA拷贝的探针，通过斑点杂交法检测甲型流感病毒RNA。为了用生物素进行标记，采用了一种通过核酸化学修饰来引入生物素的新方法。在同源DNA:DNA杂交中，经生物素处理的探针检测灵敏度小于1 pg，放射性标记的探针检测灵敏度为1.25 pg。用DNA探针与从感染甲型流感病毒（A/苏联/90/77株和A/得克萨斯/77株）的MDCK细胞中分离出的细胞质RNA进行杂交，结果显示斑点中的RNA相当于2×10⁴个细胞中存在的4.5 - 5.5 1g ID₅₀的病毒。在所有试验中，该探针在任何斑点中均不与阴性对照结合。研究结果表明，用生物素和³²P标记并用于斑点杂交以检测感染细胞中甲型流感病毒RNA的DNA探针具有相似的灵敏度和特异性。
