CAJ | 학술논문

目的：建立适用于POCT的人血清N末端B型钠尿肽原（NT-proBNP）荧光定量检测方法。方法采用免疫荧光双抗体夹心法，研制快速定量检测人血清中NT-proBNP的免疫层析检测试剂盒；通过检测线性、精密度、准确度、特异性、稳定性等指标进行试剂盒的实验室性能评估；并选取2013年2月至2014年4月广东省第二人民医院、中山大学孙逸仙纪念医院和郑州市儿童医院疑似心血管疾病患者1056例（男性605例，女性451例），采集其血清标本，通过研制试剂与参比试剂的多中心、平行比对研究进行试剂盒临床应用评估。采用相关分析、线性回归、受试者工作特征（ ROC）曲线分析、阴／阳性符合率统计学方法对数据进行统计分析。结果 NT-proBNP定量试剂报告范围为18～35000 ng／L；试剂在重复检测低、中、高3个浓度校准品的变异系数（ CV）小于15％，偏倚小于20％；标本中常见干扰物胆红素、甘油三酯、胆固醇在所测定的浓度下，测试结果的CV值可控制在15％以内，对NT-proBNP定量检测无明显影响；14个月内检测不同浓度的NT-proBNP校准品，偏倚可以控制在±20％以内，试剂有效期大于12个月；在临床样本的检测中，本试剂盒与参比试剂产品有较好的相关性（Y研制＝1.0489X参比＋121.54，R2＝0.9566，n＝1056），并且对临床血清标本定量结果的偏差无统计学意义（ Z＝0.88，P＝0.379＞0.05）；研制的NT-proBNP检测结果以参考试剂结果为标准进行比对，在2个不同cut-off 值（300和450 ng／L ）下的 ROC 曲线下面积分别为0.981和0.978。结论本研究建立的NT-proBNP免疫荧光定量层析检测方法及相应的试剂，在各项指标的评估中均达到临床检测的要求，可用于血清NT-proBNP 指标的快速检测。
목적：건립괄용우POCT적인혈청N말단B형납뇨태원（NT-proBNP）형광정량검측방법。방법채용면역형광쌍항체협심법，연제쾌속정량검측인혈청중NT-proBNP적면역층석검측시제합；통과검측선성、정밀도、준학도、특이성、은정성등지표진행시제합적실험실성능평고；병선취2013년2월지2014년4월광동성제이인민의원、중산대학손일선기념의원화정주시인동의원의사심혈관질병환자1056례（남성605례，녀성451례），채집기혈청표본，통과연제시제여삼비시제적다중심、평행비대연구진행시제합림상응용평고。채용상관분석、선성회귀、수시자공작특정（ ROC）곡선분석、음／양성부합솔통계학방법대수거진행통계분석。결과 NT-proBNP정량시제보고범위위18～35000 ng／L；시제재중복검측저、중、고3개농도교준품적변이계수（ CV）소우15％，편의소우20％；표본중상견간우물담홍소、감유삼지、담고순재소측정적농도하，측시결과적CV치가공제재15％이내，대NT-proBNP정량검측무명현영향；14개월내검측불동농도적NT-proBNP교준품，편의가이공제재±20％이내，시제유효기대우12개월；재림상양본적검측중，본시제합여삼비시제산품유교호적상관성（Y연제＝1.0489X삼비＋121.54，R2＝0.9566，n＝1056），병차대림상혈청표본정량결과적편차무통계학의의（ Z＝0.88，P＝0.379＞0.05）；연제적NT-proBNP검측결과이삼고시제결과위표준진행비대，재2개불동cut-off 치（300화450 ng／L ）하적 ROC 곡선하면적분별위0.981화0.978。결론본연구건립적NT-proBNP면역형광정량층석검측방법급상응적시제，재각항지표적평고중균체도림상검측적요구，가용우혈청NT-proBNP 지표적쾌속검측。
Objective To develop a rapid quantitative detecting assay for point-of-care testing ( POCT ) of N-terminal pro-brain natriuretic peptide ( NT-proBNP ) in serum by the fluorescence immunochromatographic technology.Methods Applying double-antibody sandwich assay to establish the quantitative NT-proBNP kit.The performance of quantitative NT-proBNP kit was evaluated by the sensitivity , specificity, accuracy, precision, stability and clinical effectiveness.It compared the research kit and conference kit by the parallel experience in the 1 056(605 males, 451 females)serum specimen collected from Guangdong Provincial People′s Hospital, Sun Yat-sen Memorial Hospital and Children′s Hospital of Zhengzhou between February 2013 to April 2014.Statistical significance of the results was assessed by correlation analysis , linear regression , receive operating characteristic ( ROC) curve analysis , negative and positive consistent.Results The report range of the NT-proBNP kit was 18-35 000 ng/L.The coefficient of variation ( CV) values for low , median and high concentration calibrators respectively were all less than 15%.Common interfering substances in human serum specimens such as bilirubin , triglyceride and cholesterol were found no significant affect on NT-proBNP antigen detection and the CV were no more than 15%.According to the results of detection for calibrators , the shelf time of the NT-proBNP diagnostic kit should be longer than 12 months.The NT-proBNP kit and reference kit had good correlation ( Y=1.048 9X developed reference +121.54, R2 =0.956 6, n=1 056) to detect the target protein through the parallel experiments and the deviation of the quantitative results of clinical serum samples showed no statistical significance (Z=0.88, P=0.379>0.05).The clinical assays of two different diagnostic kits showed good consistency based on the ROC curve evaluation which is compared by two cut-off values (300 and 450 ng/L).The areas under ROC curve were 0.981 and 0.978 respectively.Conclusions A novel NT-proBNP chromatographic quantitative immunofluorescence detection method was developed in this study .The performance evaluation data indicated that the kit is suitable for rapid detection of serum NT -proBNP.