McKeating J A, Al-Nakib W, Greenaway P J, Griffiths P D
J Virol Methods. 1985 Jul;11(3):207-16. doi: 10.1016/0166-0934(85)90109-0.
Various factors influencing the detection of human cytomegalovirus (HCMV) in infected cells by DNA-DNA hybridization have been investigated. Employing the Hind III O fragment of HCMV AD169 labelled with 32P, we found that detection sensitivity was highly influenced by the method employed for extraction of DNA from infected cells. Excision of the Hind III O fragment from the vector by restriction endonuclease digestion prior to 32P-labelling further improved the detection capability of the probe. Similarly, cytomegalovirus (CMV) DNA detection employing biotin-labelled probes and streptavidin/alkaline phosphatase in the hybridot assay was also highly dependent on the method of DNA extraction prior to hybridization. Finally, we describe an in situ assay employing a biotin-labelled probe and fluorescein-conjugated avidin to detect CMV DNA in cultured cells.
研究了多种影响通过DNA-DNA杂交在感染细胞中检测人巨细胞病毒（HCMV）的因素。使用用32P标记的HCMV AD169的Hind III O片段，我们发现检测灵敏度受从感染细胞中提取DNA的方法的高度影响。在32P标记之前通过限制性内切酶消化从载体中切除Hind III O片段进一步提高了探针的检测能力。同样，在杂交分析中使用生物素标记的探针和链霉亲和素/碱性磷酸酶进行巨细胞病毒（CMV）DNA检测也高度依赖于杂交前DNA提取的方法。最后，我们描述了一种使用生物素标记的探针和荧光素偶联抗生物素蛋白在培养细胞中检测CMV DNA的原位分析方法。
