外周血基因转录本检测中2种标本处理方法的比较  

Evaluation of two sample preparation methods in quantitative detection for gene expression in peripheral blood

作　　者：曹文俊[1] 吴华成[2] 璩斌[1] 樊绮诗[1] 

机构地区：[1]上海第二医科大学附属瑞金医院检验科,上海200025 [2]上海第二医科大学附属瑞金医院病理科,上海200025

出　　处：《上海医学检验杂志》2003年第5期268-271,共4页Shanghai Journal of Medical Laboratory Sciences

摘　　要：目的　选择合理的标本处理方法 ,提高外周血基因转录本检测的灵敏度。方法　 5 3例胃癌患者外周血标本分别以淋巴细胞分离液 (FICOLL)和红细胞裂解液 (RLS)处理后提取RNA ,以荧光定量RT PCR技术检测 β actin基因和CEA基因转录本。 结果　RLS法和FICOLL法定量检测 5 3例外周血标本的 β actin基因转录本的平均浓度分别为 1.19× 10 6和 2 .4 0× 10 5拷贝 /ml,2组测定值差异有显著性 (P <0 .0 1)。RLS法检测 5 3例中 13例为CEAmRNA阳性 ,阳性率为 2 4 .5 % ;FICOLL法检测到 5例阳性 ,阳性率 9.4 % ,RLS法的阳性检出率高于FICOLL法 (P <0 .0 5 )。结论　RLS法分离外周血有核细胞的效果优于FICOLL法 ,有利于提高基因表达检测的灵敏度。Objective To increase the sensitivity of quantitative detection of gene transcripts in peripheral blood using real time RT PCR, two specimen processing methods were compared. Methods 53 peripheral blood samples from patients with histologically confirmed gastric cancer were collected before surgery. Before RT PCR detection, blood samples were treated with both methods separately, FICOLL and red blood cell lysing solution (RLS). Mean values of β actin copy number and positivity rates of CEA mRNA were calculated and delivered for statistical analysis. Results Mean values of β actin copy number were 1.19×10 6/ml for RLS and 2.40×10 5/ml for FICOLL respectively. For the 53 patients, 13 were CEA mRNA positive (24.5%) using RLS while the positivity rate was 9.4% for FICOLL treatment. Compared with FICOLL treatment, mean value of β actin transcripts and CEA mRNA positivity rate for RLS treatment were significantly higher ( P<0.01 & P <0.05). Conclusions The efficiency of nucleared cell separation from peripheral blood through variuos sample treatment was different. We propose to use RLS be used to increase the sensitivity.

关 键 词：胃癌 Β-ACTIN基因 CEA基因 淋巴细胞分离液 红细胞裂解液 外周血基因转录本检测 

分 类 号：R735.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]