Baurin V V, Tentsov Iu Iu, Ivanova L A, Bukrinskaia A G
Vopr Virusol. 1985 Sep-Oct;30(5):540-4.
A highly sensitive method of pinpoint hybridization of nucleic acids on nitrocellulose filters using 32P-labeled pHA plasmid carrying a DNA copy of hemagglutinin gene of influenza A/Udorn/307/72 (H3N2) was developed which permitted specific detection of minimal amounts of RNA (units of pikograms) of influenza A virus with H3 serotype hemagglutinin. The method of pinpoint hybridization was used for the detection of RNA of influenza A (H3 serotype) in nasopharyngeal washings of patients with acute respiratory diseases during the influenza outbreak of February-March, 1984. In parallel, the presence of viral antigen was determined by direct immunofluorescence using H3N2 antiserum, and the diagnosis of influenza was confirmed by the clinical picture of the disease. The results indicate that the pinpoint hybridization method may be used for rapid diagnosis of influenza as a highly sensitive and specific tool.
利用携带甲型流感病毒A/Udorn/307/72(H3N2)血凝素基因DNA拷贝的32P标记的pHA质粒,开发了一种在硝酸纤维素滤膜上进行核酸精确杂交的高灵敏度方法,该方法能够特异性检测极少量具有H3血清型血凝素的甲型流感病毒RNA(皮克级单位)。在1984年2月至3月流感爆发期间,采用精确杂交方法检测急性呼吸道疾病患者鼻咽冲洗液中的甲型流感病毒(H3血清型)RNA。同时,使用H3N2抗血清通过直接免疫荧光法检测病毒抗原的存在,并根据疾病的临床表现确诊流感。结果表明,精确杂交方法作为一种高灵敏度和特异性的工具,可用于流感的快速诊断。