单克隆抗体与多克隆抗体荧光偏振免疫分析法测定环孢素A…

本文用单克隆抗荧光偏振免疫分析法（ＭＡｂ－ＦＰＬＡ）与多克隆ＦＰＬＡ法（ＰＣ－ＦＰＩＡ）测定肾移植病环孢素（ＣｓＡ）的全血浓度并进行了比较。４９例肾移植术后病人口服不同剂量ＣｓＡ期间，共监测ＣｓＡ全血谷浓度６５例次。结果表明：表ＭＡｂ－ＦＰＩＡ法观察浓度４０～６００ｎｇ／ｍｌ的范围内，ＰＣ－ＦＰＩＡ法的测定结果为１１０～１７００ｎｇ／ｍｌ，是ＭＡｂ－ＦＰＩＡ法的１．７～３．９倍。两种方法之间有良好     

荧光偏振免疫法测定环孢素A全血浓度结果分析

目的：考察不同器官移植患者环孢素谷浓度与效果之间的关系。方法：采用荧光偏振免疫法对１５７例肺、肾、骨髓移植患者进行常规ＣｓＡ血药浓度监测,并结合临床进行归纳、分析、统计。结果：发现食物、性别、肝功等均可影响ＣｓＡ的全血浓度,不同患者服用相同剂量的ＣｓＡ血药浓度差异较大。结论：在临床器官移植中,要加强血药浓度监测。     

荧光偏振免疫法测定环孢素A全血浓度结果分析

采用荧光偏振免疫法（ＦＰＩＡ）测定肾移植受者环孢素Ａ（ＣｓＡ）全血谷浓度，结果发现食物、性别、肝脏功能等均是影响ＣｓＡ全血谷浓度的重要因素；从免疫抑制效果看，术后３个月、３～６个月、６个月以上的理想稳态血浓范围分别为２００～３８０ｎｇ·ｍｌ－１、１５０～３３０ｎｇ·ｍｌ－１和１００～２５０ｎｇ·ｍｌ－１。     

特异性单克隆与非特异性多克隆抗体荧光免疫偏振法测定肾移植病人全血环孢素浓度的比较及肝功能对血药浓度的影响

目的 观察特异性单克隆抗体与非特异性多克隆抗血清荧光免疫偏振法测定肾移植病人全血环孢素浓度的差异及肝功能对血药浓度的影响。方法 对36位肾移植术后病人的84份环孢素(CsA)血样同时用特异性单克隆(MAFPIA)和非特异性多克隆抗体(PAFPIA)荧光免疫偏振分析法进行测定。对两种方法测得的数值进行统计学处理。结果MAFPIA与PAFPIA之间有较好的线性关系(相关系数为0.8947),两者的测得值具统计学显著性差异。肝功能异常的患者PAFPIA测得值显著大于MAFPIA测得值。与肝功能正常患者相比有明显差异,且两者的回归曲线参数均有明显不同。结论 对CsA血药浓度测定应采用特异性高的方法。     

单克隆荧光偏振免疫法和高效液相色谱法测定环孢素A全血浓度的相关性

目的比较单克隆荧光偏振免疫法（FPIA—m法）和高效液相色谱（HPLC）法测定环孢素A（CsA）全血浓度的相关性。方法分别采用HPLC法和FPIA—m法测定42份肾移植患者的CsA血样，测定值用t检验和线性回归方法进行分析。结果FPIA—m法与HPLC法测定值之间相关性好；回归方程为CF=78．12＋1．21CH，r=0．9893（P〈0．001）。结论FPIA—m法和HPLC法同属于特异性分析法，FPIA—m法测定值均高手HPLC法。     

两种单克隆抗体荧光免疫偏振法测定人全血中环孢素浓度的比较

目的：考察TDx与AxSYM单克隆抗体荧光免疫偏振法测定全血中环孢素A浓度结果的差异。方法：采集12名肾移植患者服药后12h内不同时间点的血样。样本分别用TDx与AxSYM进行测定。测定结果通过均数比较、Passing—Bablok回归法和Bland-Altman偏差图法进行分析。结果：TDx和AxSYM两法的测定结果相关。服药后0．5～4h的血样，两法测定结果差异无显著性。而服药6h后两法的测定结果差异有显著性，且TDx的测定结果高于AxSYM。结论：TDx与AxSYM法测定人全血中环孢素浓度的结果有差异，且与采样时间有关。     

特异性荧光偏振免疫法监测521例肾移植后环孢素A全血浓度3275次

目的 通过监测肾移植后病人环孢素A（CsA)全血浓度，提出CsA在三联免疫抑制用药方案中的理想治疗窗。方法 用特异性荧光偏振免疫法测定CsA全血浓度，对521例病人监测3275次，按术后时间及临床表现分组比较。结果 肾移植后＜1、1－3、3－6、6－12个月、1－2和＞2年的CsA全血谷浓度的理想治疗窗应分别为250－450、200－400、150－300、100－250、100－200和100－180μg/L。结论 CsA全血浓度在上述范围内，中毒反应和排异反应明显减少。     

高效液相色谱法与荧光偏振免疫法测定全血环孢菌素A浓度比较

目的探讨两种方法测定环孢菌素A浓度结果的可靠性。方法分别用高效液相法和荧光偏振免疫法测定全血中环孢菌素A浓度,比较2种方法测定结果的差异。结果2种方法的测定结果有显著性差异,FPIA法所测结果高于HPLC法。结论对环孢菌素A进行治疗药物监测时应考虑测定方法的影响。     

应用特异性荧光偏振免疫法监测406例肾移植后全血环孢素A浓度

为探讨肾移植术后环孢素A(CSA)的台理用药．应用特异性荧光偏振免疫法对406倒手术病人监测全血CSAA浓度2013次。结果表明．CSA在三联免疫抑制用药方案中的理想治疗宙范围是，术后l、1～3、3～6、6～12个月、l～2年和2年以上的全血CSA浓度分别为；256-450、200-350、180-300、105-205、100-160和90-150ug／L。在上述范围内．中毒反应和排异反应明显减少．     

荧光偏振免疫法和放射免疫法监测环孢素A血药浓度比较

荧光偏振免疫法监测全血中环孢素A血浓度与放射免疫法监测环孢素A全血和血清分析值之间都具有较好的相关性,回归方程分别为RIA=0.773 FPIA 66.8(r=0.94,P<0.05);RIA=0.378 FPIA 17.00(r=0.96,P<0.05)。FPIA/RIA比率的平均值分别是1.05±0.26和2.42±0.73。荧光偏振免疫法这种快速全自动的方法对环孢素A的体内代谢产物也同时检出,但缺乏特异性,有时应配合其他方法进行监测。     

测定环孢素全血浓度的2种方法对比及临床应用

目的对比荧光偏振免疫法(FPIA)和均相酶扩大免疫分析法(EMIT)2种测定方法监测环孢素(CsA)血药浓度的差异。方法收集98例次环孢素全血样品,同日同一患者的全血样品分别采用FPIA和EMIT法测定血药浓度。结果 EMIT和FPIA测定结果相关系数r为0.9795,EMIT法较FPIA法平均低(11±12)%(P<0.05,n=98),其中CsA血药谷值浓度(c_0)EMIT法较FPIA法平均低(14±9)%(P<0.05,n=69)。CsA血药峰值浓度(c_2)EMIT法较FPIA法平均低(2±5)%(P>0.05,n=29)。结论 2种不同方法测定全血中CsA血药浓度具有显著差异,EMIT法较FPIA法具有较高的特异性。     

特异性荧光偏振免疫法监测521例肾移植后环孢素A全血浓度3275次

目的通过监测肾移植后病人环孢素A(CsA)全血浓度 ,提出CsA在三联免疫抑制用药方案中的理想治疗窗。方法用特异性荧光偏振免疫法测定CsA全血浓度 ,对521例病人监测3275次 ,按术后时间及临床表现分组比较。结果肾移植后<1 ,、1～3、3～6、6～12个月、1～2和>2年的CsA全血谷浓度的理想治疗窗应分别为250～450、200～400、150～300、100～250、100～200和100～180μg/L。结论CsA全血浓度在上述范围内 ,中毒反应和排异反应明显减少     

同种异基因肾移植患者全血中环孢素A浓度监测结果初步分析

目的通过对该院2010年1月—2011年1月间不同肾移植患者的全血中环孢素浓度监测结果的进行初步分析,以探讨影响其浓度的可能因素,为更好地开展CsA浓度监测和临床用药提供参考。方法采用荧光偏振免疫法测定877例次肾移植患者的CsA血药浓度,分析术后时间、性别、年龄等因素与CsA血药浓度的关系。结果 CsA血药浓度随术后时间的延长而逐渐下降,各组间有统计学意义(P<0.05);男性、女性组CsA血药浓度差异无统计学意义(P>0.05);各年龄组的CsA血药浓度在同一时间段内差异无统计学意义(P>0.05)。结论 CsA血药浓度受多种因素的影响,对移植术后患者进行CsA浓度监测具有重要的临床意义。     

Comparison of cyclosporine measurement in whole blood by high-performance liquid chromatography, monoclonal fluorescence polarization immunoassay, and monoclonal enzyme-multiplied immunoassay.

Monitoring of cyclosporine concentrations in whole blood is used routinely as a guide to adjusting dose so as to achieve optimal therapeutic benefit with minimal adverse effects. In the present study, we have compared a specific high-performance liquid chromatography (HPLC) assay with a fluorescence polarization immunoassay (TDx) and an enzyme-multiplied immunoassay (Emit). Both Emit and TDx assays employ a monoclonal antibody to cyclosporin A and therefore have the potential for a high degree of specificity. Blood specimens (EDTA as anticoagulant) were obtained from 113 patients (71 renal transplants, 17 liver transplants, and 25 other categories) taking cyclosporine and analysed by all three methods. There were significant correlations between results for HPLC and Emit (Emit = 10.54 + 1.07 x HPLC; r2 = 0.82, p less than 0.001) and between results for HPLC and TDx (TDx = 9.16 + 1.42 x HPLC; r2 = 0.82, p less than 0.001). Compared to HPLC analysis, 74% and 96%, respectively, of Emit and TDx results were to the left of the line of identity. The TDx monoclonal antibody appears to have a lesser degree of specificity than that used in the Emit assay. Mean concentrations of cyclosporine measured by Emit and TDx were 17% and 51% higher, respectively, than those measured by HPLC. Because of this overestimation, we suggest that both Emit and TDx methods may find their most appropriate use in routine therapeutic monitoring of renal transplant patients in whom metabolite concentrations are less variable over time.     

荧光偏振免疫法测定血中依替米星的浓度

目的:探索应用荧光偏振免疫法测定依替米星血药浓度的实验方法,考察该方法测定依替米星的可行性.方法:使用庆大霉素荧光偏振免疫试剂盒,用TDx对依替米星进行测定.结果:本方法能够检测依替米星的血药浓度,回归曲线为Y=0.439X+0.1772,r=0.9981,在0.5～20μg·mL-1,浓度范围内线性关系良好.日内、日间标准偏差均小于10%,低、中、高不同浓度的加样回收率分别为(93.12～9,74)%,(102.41～2.73)%,(89.50～1.45)%.结论:本方法灵敏度高,操作简便,检测时间短,可以用于临床快速检测依替米星的血药浓度.     

高效液相色谱法和荧光偏振免疫法测定万古霉素血清浓度的比较

目的:比较高效液相色谱(HPLC)法与荧光偏振免疫(FPIA)法分别测定血清万古霉素浓度的结果,探讨两者的相关性。方法:收集临床检测万古霉素药物浓度的血清43份,分别用HPLC法和FPIA法进行测定,运用配对t检验,Bland-Altman分析和线性回归分析比较2种方法的测定结果。结果:HPLC法和FPIA法测定的万古霉素血清浓度具有良好的相关性, 回归方程为:Y_FPIA=1.103X_HPLC+0.831 5(R²=0.957 2); Bland-Altman评价分析2种方法一致性较好;配对t检验显示2种方法测定结果之间有显著统计学差异(P<0.000 1)。结论:相较于FPIA法,HPLC法测定不受代谢和降解产物干扰,能准确检测血清万古霉素浓度,适合应用于治疗药物监测。     

Fluorescence polarization immunoassay of mycotoxins: a review

Immunoassays are routinely used in the screening of commodities and foods for fungal toxins (mycotoxins). Demands to increase speed and lower costs have lead to continued improvements in such assays. Because many reported mycotoxins are low molecular weight (below 1 kDa), immunoassays for their detection have generally been constructed in competitive heterogeneous formats. An exception is fluorescence polarization immunoassay (FPIA), a homogeneous format that does not require the separation of bound and free labels (tracer). The potential for rapid, solution phase, immunoassays has been realized in the development of FPIA for many of the major groups of mycotoxins, including aflatoxins, fumonisins, group B trichothecenes (primarily deoxynivalenol), ochratoxin A, and zearalenone. This review describes the basic principles of FPIA and summarizes recent research in this area with regard to mycotoxins.     

Comparison of high pressure liquid chromatography and fluorescence polarization immunoassay to assess quinidine pharmacokinetics

High pressure liquid chromatography (HPLC) and fluorescence polarization immunoassay (FPIA) were compared in a quinidine pharmacokinetic study. Six healthy male subjects received single oral doses of regular release (RR) quinidine sulfate, sustained release (SR) quinidine bisulfate and the same dose of the SR product with food (SR-F). Serum was collected for 49 h after each dose and analysed for quinidine by HPLC and FPIA. Using HPLC, there were no statistically significant differences between dosing regimens with respect to area under the curve (AUC) or terminal rate constant (K). The RR dose resulted in a higher peak plasma concentration (Cp) and shorter time to peak (Tp) than either of the SR doses (p less than 0.01). Food had no apparent effect on the bioavailability of the SR product. When using FPIA, food administration was found to increase the AUC for the SR product (p less than 0.05) and all three dosage regimens resulted in a different Cp and Tp (p less than 0.001). When comparing all pharmacokinetic parameters determined by each assay, FPIA resulted in a significantly lower K (p less than 0.01). Orthogonal regression of all serum quinidine concentrations showed that FPIA = 0.926 (HPLC) + 0.06 (r = 0.971, p less than 0.001). Analysis of quinidine concentrations in different concentration ranges revealed that FPIA overestimated HPLC for concentrations less than 1 micrograms ml-1 (p less than 0.001) and underestimated HPLC for concentrations greater than 2 micrograms ml-1 (p less than 0.01). Although the use of FPIA is appropriate for quinidine therapeutic drug monitoring, HPLC is the preferred assay method for assessing pharmacokinetic parameters in single dose bioavailability studies.