人型结核杆菌全染色体DNA探针鉴别结核杆菌和其它分枝杆菌的研究

RAPID IDENTIFICATION OF MYCOBACTERIUM TUBERCULOSIS AND OTHER MYCOBACTERIA WITH MYCOBAC TERIUM TUBERCULOSIS WHOLE CHROMOSOMAL DNA PROBE

诊断结核病和其他分枝杆菌病的许多细菌免疫学传统方法敏感性低,而且烦琐费时。随着分子生物学和基因工程技术的发展,DNA 探针已成为结核病快速诊断和分枝杆菌分类、鉴定的一种敏感方法。目前全染色体 DNA 探针已能够鉴别非同源性和同源性低的菌种。本文介绍在不同杂交温度下应用 DNA 斑点杂交技术鉴别人型结核杆菌(Mycobacterium tuberculosis)

The dot-blots containing DNA isolatedfrom nonmycobacterial and mycobacterialmicroorganisms were hybridized with ^(32)P-labeled M.tuberculosis whole chromosomalDNA at the various temperatures.The probedid not cross-hybridize to DNA of non-mycobacterial microorganisms(E.coli,Plas-mid pUC19,Nocardia asteriodes),nor withDNA from all mycobacteria tested except M.bovis BCG under the higher temperature con-ditions.Microorganisms could also be dir-ectly spotted and lysed on nitrocellulose fil-ters and used for hybridization thus makingthis technique suitable for clinical diagnosis.