利用原位分子杂交结合酪酰胺放大检验军团菌性肺炎组织嗜肺军团菌的研究

发布时间：2018-06-27 09:03

本文选题：嗜肺军团菌 + 肺炎　；参考：《现代预防医学》2013年10期

【摘要】：目的利用针对嗜肺军团菌种特异性16SrRNA和军团菌致病基因—巨噬感染增强因子(mip)序列探针原位分子杂交结合酪酰胺放大技术(Tyramide signal amplification method,TSA)建立军团菌性肺炎组织中嗜肺军团菌的快速、特异性检验方法。方法设计特异性针对嗜肺军团菌种特异性16SrRNA、mip序列探针,分别采用荧光、地高辛标记探针对嗜肺军团菌性豚鼠肺炎模型支气管肺泡灌洗液、肺组织进行原位杂交结合TSA检验嗜肺军团菌。结果荧光原位杂交显示支气管肺泡灌洗液细胞中存在16SrRNA、mip序列双杂交阳性杆状细菌;地高辛标记原位杂交结合TSA显示肺组织中也存在杂交阳性杆状细菌,分布于肺泡壁与细胞内,整个检验周期小于24h。结论结果表明采用荧光、地高辛标记探针结合TSA的原位杂交能迅速准确灵敏检验出肺组织、支气管肺泡灌洗液中的嗜肺军团菌,是一种快速、特异性检验方法。
[Abstract]:Objective to establish the rapid development of Legionella pneumophila in Legionella pneumoniae by in situ hybridization with in situ hybridization and tyramide signal amplification technique using 16s rRNA specific to Legionella pneumophila and (mip) probe of Legionella pathogenicity gene-macrophage infection enhancer. Specificity test method. Methods A specific 16s rRNA mip sequence probe was designed for Legionella pneumophila strain. Fluorescent and digoxin labeled probes were used to treat bronchoalveolar lavage fluid (BALF) of Legionella pneumophila pneumonia model in guinea pigs. In situ hybridization and TSA were performed to detect Legionella pneumophila in lung tissue. Results fluorescence in situ hybridization showed that there were 16s rRNA mip positive rod bacteria in bronchoalveolar lavage fluid cells, digoxigenin-labeled in situ hybridization combined with TSA showed that there were hybridization positive rod-like bacteria in lung tissue, which distributed in alveolar wall and cell. The whole inspection period is less than 24 h. Conclusion fluorescence, digoxigenin labeled probe and in situ hybridization with TSA can quickly, accurately and sensitively detect Legionella pneumophila in lung tissue and bronchoalveolar lavage fluid. It is a rapid and specific method for detection of Legionella pneumophila in bronchoalveolar lavage fluid.
【作者单位】：西安市疾病预防控制中心;
【基金】：陕西省自然科学基金资助项目(2006C223) 西安市支撑计划资助项目(YF07154)
【分类号】：R378

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