肝螺杆菌脂多糖提取及纯化方法的建立

目的建立肝螺杆菌脂多糖（LPS）提取与纯化方法。方法采用脂多糖提取试剂盒提取肝螺杆菌LPS，并用DNaseI和RNase酶除去DNA和RNA，纯化LPS。三次测定LPS中糖、蛋白质及核酸的含量，并用SDS．PAGE电泳银染方法分析LPS纯度。结果肝螺杆菌为形态细小、Giemsa’8染色阴性的不规则螺旋型杆菌。纯化的LPS平均产率为0．202％，平均蛋白质含量为0．365g／L，平均核酸含量为0．413mg／L，三者结果的稳定性好，批间比较差异无统计学意义（P〉0．05）。SDS．PAGE电泳结果显示，LPS提取物与标准品比较，主要条带清晰，位置相同。结论建立了一种有效提取肝螺杆菌LPS的方法，该方法简便可行、提取物纯度高，核酸及蛋白质含量低，值得推广应用。

Extraction and purification of Helicobacter hepaticus LPS

Objective To establish a method for extraction and purification of Helicobacter hepaticus lipopolysaecharide （LPS）. Methods The LPS was extracted by LPS Extraction Kit.The DNA and RNA were eliminated by DNase I and RNase digestion. The lipopolysaccharide was then purified. The contents of sugar, protein and nucleic acid were measured. The purity of lipopolysaccharide was analyzed by SDS-PAGE and silver staining. Results Helicobacter hepat/cus was Giemsa＇s staining negative, small and irregular spiral bacteria. The yield of LPS was 0.202%, which contained 0.365 g/L protein, and 0.413 mg/L nucleic acid. It was stable. SDS-PAGE electrophoresis showed that LPS- extract was similar to the standard. Conclusion An effective method which can extract high purity lipopolysaccharid from Helicobacter hepaticus, with low content of nucleic acid and protein lipopolysaccharide was established, and should be widely applied.