摘要
目的比较煮沸裂解法、酚/氯仿法和试剂盒法3种DNA提取法在肺结核患者粪便标本中检测TB-DNA的应用效果。方法收集肺结核患者粪便标本,分别用煮沸裂解法、酚/氯仿法和试剂盒法提取DNA,比较其TB-DNA实时荧光聚合酶链反应(PCR)扩增的检测阳性率及扩增效率,分析不同方法的差异。结果对于痰抗酸涂片阴性患者的粪便标本,3种方法均未测到有TB-DNA扩增。痰抗酸涂片阳性患者的粪便标本,煮沸裂解法、酚/氯仿法和试剂盒法的TB-DNA检出率分别为45%、70%和80%。酚/氯仿法与试剂盒法TB-DNA的定量结果比较,存在显著性差异,试剂盒法提取扩增效果显著强于酚/氯仿法。结论试剂盒法可有效去除粪便标本中的PCR抑制物,TB-DNA检出率和定量结果明显优于煮沸裂解法和酚/氯仿法,对于无法产生痰或咳出痰的肺结核患者的实验室诊断具有较高的应用价值。
Objective To Compare three DNA extraction methods including boiling lysis method, phenol/ chloroform method,and kit method for TB DNA detection in stool specimens of patients with pulmonary tuberculosis. Methods Stool specimens of pulmonary tuberculosis patients are collected and treated separately with boiling lysis method,phenol/chloroform method and kit method. TB DNA positive detection rates and amplification efficiency of these three methods were analyzed and compared with real time PCR. Results TB DNA amplification was no de- tected in stool specimens of sputum smear negative patients. The TB DNA detection rates of these three methods were 45% ,70% and 80% ,respectively. The quantitative result of TB DNA in the kid method group was significantly higher than that in the phenol/chloroform method group. Conclusion Compared with the other two methods, kit method can more effectively remove PCR inhibitors which has a high diagnosis value for pulmonary tuberculosis patients,who can not produce or cough up sputum.
出处
《检验医学与临床》
CAS
2012年第10期1207-1209,共3页
Laboratory Medicine and Clinic
关键词
肺结核
粪便
DNA提取
实时荧光聚合酶链反应
结核分枝杆菌
Pulmonary tuberculosis
Stool
DNA extraction
Real-time fluorescence PCR
Mycobacterium tuberculosis complex
