摘要
目的对肝癌石蜡组织标本病理切片中发现的细菌进行分离鉴定. 方法 38例肝癌石蜡组织标本经聚合酶链反应(PCR)扩增螺杆菌16S rRNA,阳性标本病理切片后行石炭酸-碱性品红染色观察幽门螺杆菌(H.pylori),应用激光捕获显微切割技术(LCM)分离光镜下观察到的细菌,分离的细菌经PCR扩增螺杆菌16S rRNA,PCR产物进行测序及同源比较,阳性者再扩增H.pylori的特异基因[相对分子质量(Mr)为26×103蛋白,H.pylori种特异抗原]和相关功能基因(cagA, vacA). 结果15例肝癌石蜡组织标本中检测到螺杆菌16S rRNA,选取观察到细菌数量较多的6例用于LCM分离.分离的6例细菌样本均扩增出螺杆菌16S rRNA,经测序与H.pylori有99%～100%的同源性.4例Mr为26×103蛋白基因阳性,1例扩增出cagA基因,未扩增出vacA基因. 结论肝癌石蜡组织中经PCR检测到的H.pylori就是病理观察到的细菌.
Objective To isolate and identify the bacteria in paraffin embedded liver tissues of hepatocellular carcinoma (HCC). Methods Thirty eight paraffin-embedded tissue samples of HCC were examined by polymerase chain reaction (PCR) with Helicobacter-specific 16S rRNA primers. Histological sections of positive samples were stained by carbolic acid-basic fuchsin and the bacteria in sections were isolated by laser capture microdissection(LCM) technology. Helicobacter species 16S rRNA gene, H.pylori specific-antigen (M r 26×103) gene, cytotoxin-associated gene A(cagA), vacuolating toxin gene (vacA) were detected by PCR. Results Fifteen samples were positive in Helicobacter-specific 16S rRNA gene and 6 samples with more bacteria were selected to isolate the bacteria observed in sections by LCM. All six samples have been amplified for 16S rRNA (400 bp) by PCR, 4 samples were positive for M r 26×103, 1 positive for cagA and no positive in vacA. Three samples of 16S rRNA amplicons were sequenced and showed a 99%-100% identity to H.pylori. Conclusion H.pylori detected by PCR in liver tissues of HCC was the same one as the bacteria observed in histologic sections.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2005年第4期324-328,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目(No. 30271171)
