原位聚合酶链反应检测结核分枝杆菌L型的初步研究

Detection of -Mycobacterium tuberculosis L forms by in situ polymerase chain reaction

目的 :建立一种不需提取DNA的检测分枝结核杆菌L型的分子生物学方法。方法 :采用原位聚合酶链反应 (原位PCR)技术 ,扩增结核杆菌L型及其感染的人和小鼠血标本中的结核杆菌IS 6 110基因 ,再用免疫组化技术显色。结果 :结核杆菌L型菌株中的DNA被扩增 ,血标本中 ,可见到红细胞和淋巴细胞周围粘附大量结核杆菌DNA阳性颗粒。结论 :红细胞和淋巴细胞具有免疫粘附功能 ;原位PCR快速、简便 ,可用于检测稳定和不稳定分枝杆菌L型。

Objective:To establish a molecular biological method for detection of Mycobacterium tuberculosis (Mtb) L forms without DNA extraction.Methods: In situ PCR technique was used for amplification of Mtb IS 6110 gene in mycobacteria L forms strain and in blood specimens from patients or mice,and immunohistochemical ABC method was used for expression.Results:Amplified DNA was seen in the bacterial cells and a large number of DNA granules of Mtb clustered around lymphocytes and red blood cells.Conclusions:Red blood cells and lymphocytes have immune adherence functions, in situ PCR is a rapid,simple method of detecting stable and unstable mycobacteria L forms. [