用PCR扩增16Sr RNA基因鉴定幽门螺杆菌的细胞壁缺陷型

目的：探讨幽门螺杆菌及其稳定L型的检测与鉴定方法。方法：用抗生素诱导、滤菌器过滤、非高渗透压培养基培养获得幽门螺杆菌稳定L型纯培养物,对幽门螺杆菌及其稳定L型进行16SrRNA基因的PCR扩增及序列测定。结果：幽门螺杆菌及其稳定L型的16SrRNA基因PCR扩增产物经1%琼脂糖电泳,在紫外检测仪下可见550 bp的DNA条带,测序结果经NCB I B last分析比对,基因同源性可达100%。结论：16SrRNA基因PCR检测可用于幽门螺杆菌L型及其他形态变异的菌种鉴定。

Identification of Cell Wall Deficient Helicobacter pylori With 16SrRNA Gene PCR Amplification

Objective： To exploit effective methods for detection and identification of Helicobacter pylori and its stable cell wall-deficient form （L-form）. Methods： Stable L-form of H. pylori was induced by ceftriaxone sodium in non-hyperosmosic medium and purified by filtration. The 16SrRNA gene of L- form and normal H. pylori were amplified by PCR and analyzed with agarose gel electroforesis and DNA sequencing. Results： The PCR products were approximately 550 bp. and their sequences were verified as 100% homology with the part of H. pylori 16SrDNA gene recorded in NCBI gene bank with blasting. Conclusion： L-form H. pylori could be identified through analyzing its 16SrDNA with PCR.