十二指肠液K-ras癌基因检测在胆管癌早期诊断中的应用价值

Applied value of detection of K-ras gene mutations in duodenal juice of patients with biliary tract carcinoma in early diagnosis

目的探讨十二指肠液标本中K-ras癌基因检测对胆管癌早期诊断的价值。方法应用DNAzolR试剂提取DNA,通过聚合酶链反应-限制性片段长度多态性(polymerase chain reaction restriction fragment lengthpolymorplism,PCR-RFLP)法分析,检测30例胆管癌患者和10例良性胆道疾病患者胆汁及十二指肠液标本中上清液及沉渣的K-ras基因突变情况。结果胆管癌组胆汁标本的上清液中K-ras基因突变检测的阳性率为56.7%;相应的十二指肠液标本阳性率为43%。良性胆道疾病组仅胆汁标本检出K-ras基因突变1例,良恶性疾病组比较差异有统计学意义﹙P<0.005),胆管癌K-ras基因突变的检出率在上清液组中明显高于沉渣组,两者比较差异有统计学意义﹙P<0.005),对于胆管刷检阴性的胆管癌患者,检测胆汁中K-ras基因突变的阳性率仍可达39.3%。结论应用DNAzolR试剂能有效提取到胆汁及十二指肠液标本中的DNA,十二指肠液K-ras基因检测可能有助于胆管癌的早期诊断。

Objecive To explore the applied value of detection of K-ras gene mutations in the duodenal juice of patients with biliary tract carcinoma （BTCa） in early diagnosis. Methods DNA was extracted using DNAzol^R reagent. The situation of K-ras mutation in the supernant and sediments in the sample of bile and duodenal in 30 cases with BTCa and 10 cases with benign choledocholithiasis were detected with PCR-RFLP. Results The positive rates of assay with K-ras gene mutations in bile supernatant of patients with BTCa were 56.7%. Accordingly, the positive rates of those in duodenal juice were 43%. Only one case K-ras gene mutation was detected in benign choledochlithiasis. Statistic analysis showed there was significant difference in those gene mutation between BTCa group and benign group （P〈0.005）. Positive rate of K-ras gene mutations was higher in the supernatant than that in the sediments （P〈0.005）. For the negative of bile duct brushing in BTCa, positive rates of assay with K-ras gene mutation could still reach 39.3%. Conclusions DNA can be extracted from bile and duodenal juice with DNAzol^R reagent. Assay with K-ras gene mutations in duodenal juice may be useful for early diagnosis of biliary tract carcinoma.