中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2012年
12期
1102-1107
,共6页
方琦%黄锡荣%李凯%唐时幸%王继华
方琦%黃錫榮%李凱%唐時倖%王繼華
방기%황석영%리개%당시행%왕계화
降钙素%荧光免疫测定%床旁诊断化验信息系统
降鈣素%熒光免疫測定%床徬診斷化驗信息繫統
강개소%형광면역측정%상방진단화험신식계통
Calcitonin%Fluoroimmunoassay%Point-of-care systems
目的 利用荧光免疫层析技术,建立一种适用于床旁检验(POCT)的人血清降钙素原(PCT)快速定量检测方法.方法 采用免疫荧光双抗体夹心法(1株抗体包被于硝酸纤维素膜,另1株抗体标记荧光胶乳)研制PCT荧光免疫层析定量检测试剂盒.选取2012年1至5月广州市红十字会医院疑似细菌感染患者472例,其中,男性240例,女性232例,采集其血清标本,用本试剂盒检测PCT含量,同时通过线性、精密度、准确度、特异性、稳定性试验和与国外试剂对比分析进行方法学评价与诊断性能分析.结果 PCT定量试剂报告范围为0.1～125.0 μg/L;选取两批试剂对低、中、高3个浓度质控品重复测定20次的变异系数(CV)均小于15％,偏倚可接受(P＞0.05)；标本中常见干扰物胆红素(2.0g/L)、三酰甘油(30.0 g/L)、胆固醇(15.0 g/L)在所测定的浓度下对PCT定量检测无明显影响.14个月内检测浓度为0.5、1.0、22.0、65.0 μg/L的PCT质控品,各浓度相对偏差可以控制在±20％以内,认为试剂盒有效期＞12个月.用本试剂盒与对照产品VIDAS BRAHMS PCT定量试剂盒平行检测472份临床血清标本,两种方法有很好的相关性(YVIDAS=0.180+1.006X万孚,R2=0.988,P＜0.01),并且对临床血清标本定量结果的偏差无统计学意义(Z=-1.6,P＞0.05)；以对照产品的诊断结果为标准,万孚-PCT检测结果在3个cut-off值(0.5、2.0、10.0 μg/L)下的受试者操作特征(ROC)曲线下面积分别为0.997、0.994、0.998,P＜0.01；Kappa值分别为0.899、0.905、0.973,认为两种方法的诊断结果具有较好的一致性.结论 本研究建立了PCT荧光免疫层析快速定量检测方法,各项指标均达到临床检测的要求,可用于临床血清PCT浓度的快速检测.
目的 利用熒光免疫層析技術,建立一種適用于床徬檢驗(POCT)的人血清降鈣素原(PCT)快速定量檢測方法.方法 採用免疫熒光雙抗體夾心法(1株抗體包被于硝痠纖維素膜,另1株抗體標記熒光膠乳)研製PCT熒光免疫層析定量檢測試劑盒.選取2012年1至5月廣州市紅十字會醫院疑似細菌感染患者472例,其中,男性240例,女性232例,採集其血清標本,用本試劑盒檢測PCT含量,同時通過線性、精密度、準確度、特異性、穩定性試驗和與國外試劑對比分析進行方法學評價與診斷性能分析.結果 PCT定量試劑報告範圍為0.1～125.0 μg/L;選取兩批試劑對低、中、高3箇濃度質控品重複測定20次的變異繫數(CV)均小于15％,偏倚可接受(P＞0.05)；標本中常見榦擾物膽紅素(2.0g/L)、三酰甘油(30.0 g/L)、膽固醇(15.0 g/L)在所測定的濃度下對PCT定量檢測無明顯影響.14箇月內檢測濃度為0.5、1.0、22.0、65.0 μg/L的PCT質控品,各濃度相對偏差可以控製在±20％以內,認為試劑盒有效期＞12箇月.用本試劑盒與對照產品VIDAS BRAHMS PCT定量試劑盒平行檢測472份臨床血清標本,兩種方法有很好的相關性(YVIDAS=0.180+1.006X萬孚,R2=0.988,P＜0.01),併且對臨床血清標本定量結果的偏差無統計學意義(Z=-1.6,P＞0.05)；以對照產品的診斷結果為標準,萬孚-PCT檢測結果在3箇cut-off值(0.5、2.0、10.0 μg/L)下的受試者操作特徵(ROC)麯線下麵積分彆為0.997、0.994、0.998,P＜0.01；Kappa值分彆為0.899、0.905、0.973,認為兩種方法的診斷結果具有較好的一緻性.結論 本研究建立瞭PCT熒光免疫層析快速定量檢測方法,各項指標均達到臨床檢測的要求,可用于臨床血清PCT濃度的快速檢測.
목적 이용형광면역층석기술,건립일충괄용우상방검험(POCT)적인혈청강개소원(PCT)쾌속정량검측방법.방법 채용면역형광쌍항체협심법(1주항체포피우초산섬유소막,령1주항체표기형광효유)연제PCT형광면역층석정량검측시제합.선취2012년1지5월엄주시홍십자회의원의사세균감염환자472례,기중,남성240례,녀성232례,채집기혈청표본,용본시제합검측PCT함량,동시통과선성、정밀도、준학도、특이성、은정성시험화여국외시제대비분석진행방법학평개여진단성능분석.결과 PCT정량시제보고범위위0.1～125.0 μg/L;선취량비시제대저、중、고3개농도질공품중복측정20차적변이계수(CV)균소우15％,편의가접수(P＞0.05)；표본중상견간우물담홍소(2.0g/L)、삼선감유(30.0 g/L)、담고순(15.0 g/L)재소측정적농도하대PCT정량검측무명현영향.14개월내검측농도위0.5、1.0、22.0、65.0 μg/L적PCT질공품,각농도상대편차가이공제재±20％이내,인위시제합유효기＞12개월.용본시제합여대조산품VIDAS BRAHMS PCT정량시제합평행검측472빈림상혈청표본,량충방법유흔호적상관성(YVIDAS=0.180+1.006X만부,R2=0.988,P＜0.01),병차대림상혈청표본정량결과적편차무통계학의의(Z=-1.6,P＞0.05)；이대조산품적진단결과위표준,만부-PCT검측결과재3개cut-off치(0.5、2.0、10.0 μg/L)하적수시자조작특정(ROC)곡선하면적분별위0.997、0.994、0.998,P＜0.01；Kappa치분별위0.899、0.905、0.973,인위량충방법적진단결과구유교호적일치성.결론 본연구건립료PCT형광면역층석쾌속정량검측방법,각항지표균체도림상검측적요구,가용우림상혈청PCT농도적쾌속검측.
Objective To develop a quick quantitative detecting method for point of care testing (POCT) of human serum procalcitonin (PCT) by fluorescence immunochromatographic technology.Methods Applying a double-antibody sandwich immunofluorescent assay (one antibody coated on the nitrocellulose membrane and the other antibody labeled with fluorescent micropaticles) to develop a PCT quantitative detecting kit by immunochromatography technology.The kit was used to test PCT in 472 serum samples from suspected bacterial infection patients of Guangzhou Red Cross Hospital,including 240 male and 232 female patients.The methodology and diagnostic performance were evaluated in the aspects of linearity,precision,accuracy,specificity,stability experiments and comparison with foreign PCT detecting kits.Results The report range of the PCT quantitative diagnostic kit was 0.1-125.0 μg/L The coefficient of variation (CV)values of repeat 20 tests for low,median,and high concentration control samples respectively were all less than 15％ and bias can be acceptable (P ＞ 0.05).Common interfering substances in human serum specimens such as bilirubin (2.0 g/L),triglyceride (30.0 g/L) and cholesterol (15.0 g/L) were found no significant affect on quantitative detection of PCT.The shelf time of the PCT diagnostic kit should be longer than 12 months as the relative deviation of detected concentrations of 0.5,1.0,22.0,65.0 μg/L PCTcontrol sample can be controlled less than 20％ within 14 months.Considering VIDAS BRAHMS PCT to be the standard quantitative test for PCT,472 serum samples were detected by both our kit and the control VIDAS BRAHMS PCT kit simultaneously,which showed high correlation (YVIDAS =0.180 + 1.006Xwondfo,R2 =0.988,P ＜ 0.01) and low deviation (Z =-1.6,P ＞ 0.05) without statistic significance between two methods.And the results of these two diagnostic kits showed good consistency as the area under curve of the receiver operating characteristic (ROC) of Wondfo-PCT at the three cut-off values (0.5,2.0,10.0 μg/L)were 0.997,0.994,0.998 respectively,P ＜ 0.01,using diagnostic result of the control product as standard.Kappa values were 0.899,0.905,0.973 respectively.Conclusions The method of quantitative detection of PCT by fluorescence immunochromatography for POCT was established in this study.All the observed indicators reached the clinical diagnostic requirements and can be applied for the quick detection of clinical human serum PCT.
