Overcome false - positive in detection of tumor microme- tastasis by reverse transcriptase - polymerase chain reaction

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Overcome false - positive in detection of tumor microme- tastasis by reverse transcriptase - polymerase chain reaction

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- Overcome false - positive in detection of tumor microme- tastasis by reverse transcriptase - polymerase chain reaction
- Journal of Chongqing Medical University Issue 2, (1998)
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  CLC： R73-37
- Published：1998，
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[1]葛明建,李良彬,张玉洪,王梅,黄天禄.克服RT-PCR法检测肿瘤微转移的假阳性[J].重庆医科大学学报,1998(02):153-156.

Overcome false - positive in detection of tumor microme- tastasis by reverse transcriptase - polymerase chain reaction. [J]. Journal of Chongqing Medical University (2).(1998)
[1]葛明建,李良彬,张玉洪,王梅,黄天禄.克服RT-PCR法检测肿瘤微转移的假阳性[J].重庆医科大学学报,1998(02):153-156. DOI：

Overcome false - positive in detection of tumor microme- tastasis by reverse transcriptase - polymerase chain reaction. [J]. Journal of Chongqing Medical University (2).(1998) DOI：

摘要

为探讨克服ＲＴ－ＰＣＲ法检测恶性肿瘤微转移时出现假阳性的方法，作者运用该法检测了４０例非小细胞肺癌和２０例食管癌患者病理证实有转移的淋巴结中角蛋白１９（ＣＫ１９）基因的表达，同时还分别运用套式ＲＴ－ＰＣＥ或ＲＴ－ＰＣＲ法检测了１０例良性肺和食管肿瘤患者正常淋巴结中ＣＫ１９基因的表达。结果发现设立分别以未经逆转录的ＲＮＡ和基因组ＤＮＡ为模板的对照可观察到是否存在ＣＫ１９假基因的干扰所造成的假阳性，而在逆转录前用脱氧核糖核酸酶（ＤＮａｓｅ）消化ＲＮＡ可消除这种假阳性。运用套式ＲＴ－ＰＣＥ法测定ＤＮａｓｅ消化后的正常淋巴结ＲＮＡ可发现ＣＫ１９基因非法转录所致假阳性，调整方法灵敏度，使之刚好能区分正常和转移淋巴结是解决此类假阳性的合适办法。

Abstract

his study aimed to overcome false - positive in detection of tumor micrometastases by reverse transcriptase - polymerase chain reaction (RT - PCR)

Using this method

the authors amplified cytokeratin 19(CK19) mRNA in histologically involved regional lymph nodes of 40 patients with non - small - cell - lung - carcinoma and 20 with esophgus cancer. In addition

they amplified CK19 mRNA in DNase - pretreated normal nodes of 10 . respectively. They could find out the interference of CK19 pseudogene by performing nontranscribed RNA and genomic DNA controls. This influence could be eliminatde by treating the RNA samples with DNase before complement DNA were synthesized. Using nested RT- PCR method

they could find the presences. of illegitimate transcription of CK19 gene in DNase - pretreated normal nodes.It is a suitable measure to remove this false - positive result that regulate the sensitivity of method to distinguish the normal and involved lymph nodes exactly.

关键词

恶性肿瘤转移诊断淋巴结角蛋白聚合酶链反应

Keywords

Malignancymetastasisdiagnosislymph nodeKeratinPolymerase chain reaction

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Related Institution

Department of Thoracic Surgery, College of Clinical Medicine, Chongqing Medical University

,重庆医科大学临床学院胸心外科

重庆医科大学临床学院检验科,重庆医科大学临床学院检验科重庆　400016

Department of Cardiac-thoracic Diseases,the First Affiliated Hospital,Chongqing Medical University

重庆医科大学附属第一医院胸心外科

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