Viral Diagnosis by in situ Hybridization: Description of a Rapid Simplified Colorimetric Method
Viral Diagnosis by in situ Hybridization: Description of a Rapid Simplified Colorimetric Method
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原位杂交病毒诊断：快速简化比色法的描述
DOI：
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发表时间：
1986
影响因子：
5.6
通讯作者：
D. Myerson
中科院分区：
文献类型：
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作者：
E. Unger;L. Budgeon;D. Brigati;D. Myerson
已经开发出一种改进的比色原位杂交方法，用于在标准福尔马林固定、石蜡包埋的组织切片中诊断病毒感染。该方法采用与生物素标记的 DNA 探针杂交 2 小时，然后用亲和素-碱性磷酸酶复合物进行直接比色检测。切割组织切片后 8 小时内获得视觉结果。通过尸检或活检，已在受感染的肺组织中实现了巨细胞病毒和腺病毒遗传信息的特定组织学定位。组织和探针 DNA 在高温 (100-105°C) 下同时变性导致信号增强。我们建议，可能需要这些变性条件来使福尔马林交联的组织 DNA 更充分地变性，并有利于探针渗透到组织中。比较用于诊断 CMV 感染的杂交和病毒培养结果表明，在临床情况下，杂交将比病毒培养更快地特异性诊断有效病毒感染，但敏感性会有所损失。比色原位 DNA 杂交为外科病理学家提供了一种强大的新技术，在病毒病原体的诊断中提供了免疫细胞化学和电子显微镜的替代方法。
An improved method of colorimetric in situ hybridization for the diagnosis of viral infections in standard formalinfixed, paraffin-embedded tissue sections has been developed. This method employs a 2-hour hybridization with biotin-labeled DNA probes followed by direct colorimetric detection with avidin-alkaline phosphatase complexes. Visual results are obtained within 8 h of cutting the tissue section. Specific histologic localization of cytomegalovirus and adenovirus genetic information has been achieved in infected lung tissues from autopsy or biopsy. Simultaneous denaturation of tissue and probe DNA at elevated temperature (100-105°C) resulted in increased signal. It is our suggestion that these denaturing conditions may be required to denature more fully formalin cross-linked tissue DNA and favor penetrance of probe into the tissues. Comparison of the results of hybridization and viral culture for the diagnosis of CMV infections suggest that in clinical situations hybridization will allow specific diagnosis of productive viral infection more rapidly than viral culture with some loss in sensitivity. Colorimetric in situ DNA hybridization offers the surgical pathologist a powerful new technique that provides an alternative to immunocytochemistry and electron microscopy in the diagnosis of viral pathogens.