细胞壁缺陷幽门螺杆菌尿素酶基因活性检测  

Detection of urease and urease gene activities in Helibacter pylori with cell wall deficiency

作　　者：王菲[1] 陈峥宏[1] 康沛萍[1] 

机构地区：[1]贵州省贵阳医学院微生物学教研室,贵阳550004

出　　处：《中国公共卫生》2010年第7期936-937,共2页Chinese Journal of Public Health

基　　金：贵州省科技厅基金(黔科合J[2008]2165)

摘　　要：目的了解细胞壁缺陷变异对幽门螺杆菌尿素酶活性及其基因的影响。方法用抗生素诱导幽门螺杆菌稳定L型(HPL)并分离培养,检测幽门螺杆菌及其稳定L型的尿素酶活性,并对HPL型及其亲代细菌型进行尿素酶结构基因UreA和UreB基因的PCR检测。结果幽门螺杆菌可在头孢曲松钠作用下发生细胞壁缺陷变异,经传代培养可获得稳定L型的纯培养物;幽门螺杆菌尿素酶试验阳性,稳定L型的尿素酶试验则为阴性;L型及亲代细菌型UreA和UreB基因的PCR检测均可见分子量分别为400和200bp的扩增产物。结论幽门螺杆菌在抗生素作用下可发生细胞壁缺陷变异成为L型;幽门螺杆菌L型仍然保留UreA和UreB基因。Objective To probe the effect of cell wall deficiency on urease and urease gene activity in Heliobater pylori.Methods The stable L-form of Heliobater pylori（H pylori） were induced by ceftriaxone sodium and isolated with filtration and then subcultured in non-high osmotic medium without antibiotic.H pylori and its L-forms were identified by PCR of 16srDNA.The UreA gene and UreB gene was amplified with the specific PCR primer and the PCR products were detected with agarose gel electroforesis.Results L-form H pylori could be induced by ceftriaxone sodium in non-high osmotic liquid medium.The shape of L-form H pylori were spherical or elliptical.The single or paired L-form H pyloric arranged in chain or in pile.The stable L-forms could pass through the filter with 0.22 μm aperture and be subcultured in the non-high osmotic liquid medium without antibiotic or other inducer.The 16SrRNA gene fragment could be amplified from the stable L-form H pylori.The stable L-form derived from H pylori had UreA gene and UreB gene with the same nucleotide sequence as that of their parental bacteria.Conclusion The cell wall deficient mutation would happen when the H pylori is treated with ceftriaxone sodium,and the pure culture of stable L-form strains could be isolated with filtration and then subcultured in nonhigh osmotic medium without antibiotic.The stable L-form strains derived from H pylori have UreA and UreB gene with the same nucleotide sequence as that of their parental bacteria.

关 键 词：幽门螺杆菌 尿素酶基因 聚合酶链反应 细胞壁缺陷 

分 类 号：R994.1[医药卫生—毒理学]