王琼,陈峥宏,杨杰,潘科,熊妍,印琳,郭长城
WANG Qiong,CHEN Zhenghong,YANG Jie,PAN Ke,XIONG Yan,YIN Lin,GUO Changcheng

• 1:贵州医科大学微生物学教研室
• 2:贵州医科大学
• 3:贵州医科大学附院消化内科
• 4:黔南州人民医院消化内科

摘要(Abstract)：

目的:对球形幽门螺杆菌(Helicobacter.Pylori,H.pylori)进行鉴定,探讨其临床意义。方法:胃镜下确诊的胃炎及消化性胃溃疡患者的胃黏膜组织标本进行H.pylori分离培养、尿素酶检测,对革兰阴性(G~-)弯曲杆菌和不典型球菌进行H.pylori特异性PCR鉴定,并对特异性PCR阳性的不典型球菌以H.pylori 16S r DNA通用引物扩增测序。结果:共获得80株G~-弯曲杆菌和5株不典型球菌,在传代培养中有24株弯曲杆菌发生球形变异;G~-弯曲杆菌的组织及细菌培养物尿素酶试验均为阳性,G~-不典型球菌的组织及细菌培养物为阴性,返祖为G~-弯曲杆菌后则为阳性;85株菌经特异性引物鉴定均为H.pylori阳性,G~-不典型球菌16S rDNA序列与数据库中H.pylori 16S rRNA基因序列碱基一致性均达99%,鉴定为H.pylori。结论:H.pylori可发生球形变异,这可能会造成临床实验室微生物学诊断的假阴性。
Objective: To identify Helicobacter pylori coccid forms isolated from gastric mucosal specimens and to explore the clinical significance. Methods: Mucosal specimens of gastritis and digestibility gastric ulcer patients confirmed by gastroscope were cut into small pieces and cultured for bacterial growth. The rapid urease test was performed on gastric mucosal homogenate and bacterial cultures.Gram negative spiral bacteria and atypical spherical coccus were identified by PCR using specific primers. Then 16 S rDNA of these atypical spherical strains were amplified by universal primers,and the PCR products were sequenced. Results: Eighty strains of Gram negative campylobacter and 5 atypical spherical strains were isolated,and 24 of the campylobacter transformed into coccid form during subculture. The urease tests of the 80 typical Gram negative campylobacter were positive both in the gastric mucosa tissue and in pure cultures. However,the urease tests of mucosal tissue from which the 5strains of atypical spherical strains isolated were negative,as well as the pure culture of the spherical strains. But the urease tests of the subculture of such spherical strains changed to positive when the bacteria reverted to spiral form. Those 85 strains were all positive in PCR by using specific primers.Also the 16 S rDNA sequences of spherical strains were 99 % identical to the 16 S rRNA gene sequence of H. pylori in the NCBI database,and were identified as H. pylori. Conclusion: H. pylori can easily transform to coccid form,which could result in false negative microbiological diagnosis.

关键词(KeyWords)： 胃溃疡;胃炎;胃黏膜;螺杆菌,幽门;球形;鉴定
gastric ulcer;gastritis;gastric mucosa;helicobacter,pylorus;sphere;identification

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81460314);; 贵州省科技厅-贵阳医学院联合基金(黔科合LH2014-7075)

作者(Author): 王琼,陈峥宏,杨杰,潘科,熊妍,印琳,郭长城
WANG Qiong,CHEN Zhenghong,YANG Jie,PAN Ke,XIONG Yan,YIN Lin,GUO Changcheng

DOI: 10.19367/j.cnki.1000-2707.2017.01.007

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