银染巢式RT-PCR检测胃癌淋巴结微转移方法  

Studies on the Method for Detecting Gastric Cancer Micrometastases in Axillary Lymph Nodes by Nest RT PCR and Silver Staining

作　　者：陈瑞川[1,2,3] 陈路芸[1,2,3] 乔玉欢 邱达泰 杨善民[1,2,3] 汪德耀 

机构地区：[1]厦门大学抗癌研究中心 [2]厦门大学生物学系 [3]福建医科大学医疗系

出　　处：《厦门大学学报（自然科学版）》1999年第2期310-314,共5页Journal of Xiamen University：Natural Science

基　　金：福建省卫生厅基金;厦门大学肿瘤细胞工程国家教委开放实验开放基金

摘　　要：以Ｋｅｒａｔｉｎ１９ｃＤＮＡ的套式引物建立了巢式ＲＴ－ＰＣＲ扩增Ｋｅｒａｔｉｎ１９ｍＲＮＡ的体系及ＰＡＧＥ－银染检测扩增产物的方法；对扩增体系进行优化后，分析了扩增特异性及检测敏感性，并对临床样品作了初步检测．结果表明：该扩增体系具有较好的特异性，其敏感性可达１０－６μｇＲＮＡ，即可从１０５个淋巴细胞中检出１个胃癌细胞；对临床样品检测的结果与病理检查结果相符．显示该法具有较好的可靠性．Keratin 19 is one of cytokeratin genes which is expressed in gastric tissue and gastric carcinoma, but unexpresed in lymph tissue. With this defferent expression pattern, the gastric cancer micrometases in gastric axillary lymph nodes can be estimated by RT PCR method for detecting the Keratin 19 mRNA in total RNA from the nodes. Described here is a RT PCR system for detecting Keratin 19 mRNA with nest primers and a PAGE silver staining method for showing amplification products. The test results showed that this system possessed a good specificity and high sensitivity of detecting 10 -6 μg RNA or one gastric cancer cell from 10 5 lymph cells. The detection results with RT PCR performed on clinical gastric axillary lymph nodes were correspond to the results of pathological examination, which suggested a good application prospects in clinical.

关 键 词：胃癌转移 RT-PCR 银染 检测 淋巴结微转移 

分 类 号：R735.204[医药卫生—肿瘤]