摘要
目的 建立应用显微切割技术鉴定病毒核内包涵体的新方法 .方法 通过光学显微镜直视下用显微操纵系统进行显微切割技术结合 DNA提取和 PCR方法 ,对在组织切片上获取病毒核内包涵体的来源进行分子生物学鉴定 .结果 在福尔马林固定石蜡包埋的腮腺组织切片上根据形态学特征疑为人巨细胞病毒 (HCMV)感染的细胞 ,利用显微切割技术将核内包涵体病变与其周围染色质分离 ,从收集的包涵体得到用于 PCR扩增的 DNA模板 ,经双重 PCR扩增出预期的1 4 9bp片段 ,从而鉴定出该包涵体为 HCMV包涵体 .结论 显微切割技术可用于病毒核内包涵体的鉴定 ,为病毒包涵体病变的分子病理学研究建立了一种新的方法 .
AIM To establish a new protocol for the determination of viral intranuclear inclusions with microdissection technique. METHODS Microdissection technique with micromanipulator under microscope, as well as subsequent DNA extraction and polymerase chain reaction amplification methods, was employed. RESULTS Intranuclear inclusions with morphologic features of human cytomegarovirus in formalin fixed and paraffin embedded salivary gland tissue sections were separated from their surroundings by microdissection technique. Template DNA used for PCR was obtained from the dissected inclusions and the expected 149 bp fragments were amplified by PCR. CONCLUSION Microdissection technique can be used for the determination of viral intranuclear inclusions. A new protocol for the research of viral inclusion in molecular pathology has been established.
出处
《第四军医大学学报》
2000年第1期6-8,共3页
Journal of the Fourth Military Medical University
基金
全军医药卫生科研基金!( 98D0 46)
陕西省自然科学基金!( 98M3 8
关键词
显微切割
病毒
包涵体
microdissection
virus
intranuclear inclusions
