中国临床保健杂志
中國臨床保健雜誌
중국림상보건잡지
CHINESE JOURNAL OF CLINICAL HEALTHCARE
2014年
1期
63-67
,共5页
程民%沈国栋%卞庚%徐婷娟%徐维平%沈干%胡世莲
程民%瀋國棟%卞庚%徐婷娟%徐維平%瀋榦%鬍世蓮
정민%침국동%변경%서정연%서유평%침간%호세련
乳腺肿瘤%试剂盒,诊断%RNA,信使%逆转录聚合酶链反应
乳腺腫瘤%試劑盒,診斷%RNA,信使%逆轉錄聚閤酶鏈反應
유선종류%시제합,진단%RNA,신사%역전록취합매련반응
Breast neoplasms%Reagent kits,diagnostic%RNA,messenger%Reverse transcriptase polymerase chain reaction
目的建立人乳腺癌特异性基因1(BCSG1)mRNA实时荧光定量逆转录-聚合酶链反应(RTPCR)检测试剂盒,并评价其在乳腺癌循环肿瘤细胞检测中的应用价值。方法优化试剂盒中各组分的浓度、确定试剂盒的稳定性、灵敏度及特异性,并检测外周血样品中BCSG1mRNA的表达水平。结果优化了试剂盒中的引物、探针,成功制备了定值标准品,通过设立阴、阳性质控品的方法,解决了试剂盒的特异性、灵敏性、准确性及质量控制等关键技术问题,特异度及准确性均为95%以上,灵敏度为100拷贝/ml(全血)。外周血标本中BCSG1mRNA的检测结果为,12例临床确诊已转移的乳腺癌患者均为阳性,21例临床未确诊转移的乳腺癌患者中12例为阳性,其余9例为阴性,15例乳腺良性疾病患者、10例正常人均为阴性。结论建立的BCSG1mRNA实时荧光定量RT-PCR检测试剂盒具有很高的临床应用价值,可用于检测外周血样品中BCSG1mRNA的表达量,检测结果可作为乳腺癌患者诊疗过程中的重要监测指标。
目的建立人乳腺癌特異性基因1(BCSG1)mRNA實時熒光定量逆轉錄-聚閤酶鏈反應(RTPCR)檢測試劑盒,併評價其在乳腺癌循環腫瘤細胞檢測中的應用價值。方法優化試劑盒中各組分的濃度、確定試劑盒的穩定性、靈敏度及特異性,併檢測外週血樣品中BCSG1mRNA的錶達水平。結果優化瞭試劑盒中的引物、探針,成功製備瞭定值標準品,通過設立陰、暘性質控品的方法,解決瞭試劑盒的特異性、靈敏性、準確性及質量控製等關鍵技術問題,特異度及準確性均為95%以上,靈敏度為100拷貝/ml(全血)。外週血標本中BCSG1mRNA的檢測結果為,12例臨床確診已轉移的乳腺癌患者均為暘性,21例臨床未確診轉移的乳腺癌患者中12例為暘性,其餘9例為陰性,15例乳腺良性疾病患者、10例正常人均為陰性。結論建立的BCSG1mRNA實時熒光定量RT-PCR檢測試劑盒具有很高的臨床應用價值,可用于檢測外週血樣品中BCSG1mRNA的錶達量,檢測結果可作為乳腺癌患者診療過程中的重要鑑測指標。
목적건립인유선암특이성기인1(BCSG1)mRNA실시형광정량역전록-취합매련반응(RTPCR)검측시제합,병평개기재유선암순배종류세포검측중적응용개치。방법우화시제합중각조분적농도、학정시제합적은정성、령민도급특이성,병검측외주혈양품중BCSG1mRNA적표체수평。결과우화료시제합중적인물、탐침,성공제비료정치표준품,통과설립음、양성질공품적방법,해결료시제합적특이성、령민성、준학성급질량공제등관건기술문제,특이도급준학성균위95%이상,령민도위100고패/ml(전혈)。외주혈표본중BCSG1mRNA적검측결과위,12례림상학진이전이적유선암환자균위양성,21례림상미학진전이적유선암환자중12례위양성,기여9례위음성,15례유선량성질병환자、10례정상인균위음성。결론건립적BCSG1mRNA실시형광정량RT-PCR검측시제합구유흔고적림상응용개치,가용우검측외주혈양품중BCSG1mRNA적표체량,검측결과가작위유선암환자진료과정중적중요감측지표。
Objective This study was to establish a quantitative method for human breast cancer-specific gene 1(Breast cancer-specific Gene 1,BCSG1)mRNA by real-time RT-PCR.Its application was further evaluated by detecting circulation tumor cells in the peripheral blood of breast cancer patients .Method The concentration of each component in the detection kit was optimized;the stability,sensitivity and specificity were determined .The expression level of BCSG1 mRNA was detected in the samples of peripheral blood .Results Optimized primers and probes were obtained,and setting standards were successfully prepared .Through establishing the positive and negative control ma-terials,the specificity,sensitivity,accuracy and quality control were determined for the detection kit .The specificity and accuracy were both more than 95%,and the sensitivity was of 100 copies/ml( whole peripheral blood ) .The detec-tion results of BCSG1 mRNA in peripheral blood samples showed that all the samples were positive for 12 breast canc-er patients clinically diagnosed with metastasis;and 12 samples were positive and other 9 samples were negative among 21 breast cancer patients without clinically diagnosed with metastasis or not ;all the samples were negative for 15 pa-tients with benign breast disease;and all the samples were negative for 10 healthy control .Conclusion The quantita-tive RT-PCR assay for BCSG1 mRNA could be used for the detection of circulating tumor cells in peripheral blood of breast cancer patients ,which would be valuable for the diagnosis and clinical-decision making for breast cancer patients .