提高胃黏膜活检标本幽门螺杆菌分离率和复苏率的研究  

STUDY ON THE IMPROVEMENT OF ISOLATION AND RECOVERY RATE OF HELICOBACTER PYLORI FROM GASTRIC BIOPSY SPECIMENS

作　　者：王娜[1] 张牧霞[1] 赵萌[1] 张瑶[1] 

机构地区：[1]河北医科大学第三医院实验中心,河北石家庄050051

出　　处：《河北医科大学学报》2010年第6期679-681,共3页Journal of Hebei Medical University

基　　金：河北省自然科学基金项目(C2006000917);河北省卫生厅科研基金项目(08-144)

摘　　要：目的探索提高胃黏膜活检标本幽门螺杆菌(helicobacter pylori,H.pylori)分离率和复苏率的培养流程。方法应用145例尿素酶阳性、布氏肉汤培养基运送的胃黏膜活检标本。其中75例随机接种于布氏肉汤固体培养基、哥伦比亚琼脂培养基和脑心浸液固体培养基中,在37℃微需氧条件下培养7d,比较各培养基H.pylori阳性率;H.pylori经30%甘油布氏肉汤、30%甘油脑心浸液保存液-70℃冻存、哥伦比亚琼脂培养基复苏后,观察不同保存液对H.pylori复苏率的影响。另选70例标本,通过初次培养、2次及3次接种的方法增殖H.pylori,-70℃冻存后再复苏,观察接种次数对H.pylori复苏率的影响。结果哥伦比亚琼脂培养基的H.pylori阳性率明显高于布氏肉汤和脑心浸液固体培养基(P<0.01,P<0.05);H.pylori的复苏率在30%甘油布氏肉汤冻存液中高于30%甘油脑心浸液(P<0.05);经2、3次接种后冻存的H.pylori复苏率显著提高。结论通过布氏肉汤培养基运送、哥伦比亚琼脂培养基接种和多次传代培养、30%布氏肉汤培养基保存、复苏时用哥伦比亚培养基接种培养的程序,明显提高临床H.pylori菌株的分离培养阳性率和复苏率。Objective To explore a procedure which can improve the isolation rate and recovery rate of helicobacter pylori （H. pylori）from gastric biopsy specimens. Methods A total of 145 gastric biopsy specimens positve for urease test were used in this study. Seventy - five specimens were collected in the Brucella broth transport medium and plated randomly on Columbia agar, Brucella agar and brain heart infusion agar plates. The plates were incubated at 37~C under microaerobic conditions for 7 days. The H. pylori isolation rate was evaluated. H. pylori strains were harvested and stored at -70~C both in Brucella broth containing 30% glycerol and brain heart infusion containing 30% glycerol, and then thawed and cultured on Columbia agar plate to observe the recovery rate of H. pylori in different storage solution. Other 70 specimens were carried out primary culture, secondary subculture and the third subculture on Columbia agar plate. H. pylori strains were collected and frozen at - 70~C after each culture, and resuscitated again to determine the effect of subculture times on the recovery rate of H. pylori. Results The isolation rate of H. pylori on Columbia agar plate was significantly higher than that on Brucella broth agar and brain heart infusion agar plates . The recovery rate of H. pylori storaged in Brucella broth containing 30% glycerol was higher than that in brain heart infusion containing 30% glycerol. Frozen H. pylori strains from the secondary subculture and third subculture have higher recovery rate than that of primary culture. Conclusion By means of transporting H. pylori in Brueella broth, plating H. pylori on Columbia agar plate, proceeding subculture and storing H. pylori in Brucella broth containing 30% glycerol, will improve H. pylori isolation and get high recovery rate from frozen strain.

关 键 词：幽门螺杆菌 培养技术 胃黏膜 

分 类 号：R573[医药卫生—消化系统]