摘要
目的:评价实验室长期多次传代对幽门螺杆菌(Helicobacter pylori,H.pylori)标准菌株26695 cagA启动子基因稳定性的影响.方法:PCR方法扩增54个不同批次保存的经过多次传代的H.pylori 26695克隆子的cagA启动子,并进行测序,使用Vector NTI Suite 6软件比对序列.结果:54个H.pylori 26695克隆子的cagA启动子序列有22个(40.74%)与NCBI公布的序列完全一致,32个(59.26%)在不同位点发生碱基置换、缺失、插入等突变,其中第17位碱基有5个(15.63%)发生T→C突变,第199位碱基有14个(43.75%)发生A→C的变异,在159-160重复碱基A处有12个(37.5%)发生缺失1个或者2个A,还有一个多出2个A.结论:长期多次传代会导致H.pylori cagA启动子突变,实验室使用标准菌株要注意传代记录,建立不同等级实验室质控,根据使用标准菌株研究内容的水平决定实验室质控的等级.
AIM：To assess the genetic stability of the cagA promoter in a laboratory strain（26695） of Helicobacter pylori（H.pylori） after long-term multiple subcultures. METHODS：The promoter regions in the cagA genes of 54 subclones of the H.pylori 26695 strain,which has been subcultured many times for a long term,were amplified.PCR products were sequenced and analyzed using Vector NTI Suite 6 software. RESULTS：Twenty-two（40.74%） strains had identical sequence with the H.pylori 26695 strain, while 32 strains（59.26%） had mutations（including substitutions,deletions and insertions） at differ- ent sites.Mutations in T17C and A199C were found in 5（15.63%） and 14（43.75%） strains, respectively.Both single nucleotides or dinucleotides at positions 159 and 160 were found missing in 12（37.5%） strains,while one strain had two A insertions after position 160 in the promoter of cagA. CONCLUSION：Subcultures can lead to mutations in the cagA promoter in H.pylori 26695.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第32期3365-3369,共5页
World Chinese Journal of Digestology
