三种他克莫司测定试剂盒测定移植病人全血中他克莫司浓度的比较

目的:比较国产他克莫司测定试剂盒(微粒子酶联免疫法检测法,MEIA)、进口他克莫司测定试剂盒(微粒子酶联免疫法检测法,MEIA)以及进口他克莫司测定试剂盒(化学发光微粒子免疫法,CMIA)在检测移植病人全血中他克莫司血药浓度时的异同与相关性。方法:用国产和进口他克莫司测定试剂盒(微粒子酶联免疫法检测法,MEIA)分别测定147例移植后服用他克莫司的病人全血样品,用进口他克莫司测定试剂盒(化学发光微粒子免疫法,CMIA)测定其中49例样品,对检测结果进行统计学分析,考察三种试剂盒的等效性与相关性。结果:三种方法中任意两种方法的检测结果用配对t检验进行检测,均无显著差异,用Ex-cell进行相关性分析,其相关系数均大于0.9,相关性好。结论:三种试剂盒均可用于他克莫司的血药浓度检测,且相关性高。     

血站用部分ELISA试剂的稳定性分析与评价

目的探讨本站使用的乙型肝炎病毒表面抗原、丙型肝炎病毒抗体ELISA试剂的批内和批间变异,以评价ELISA试剂在本站使用过程中的稳定性。方法室内质控品与血液检测标本在相同的检测条件下进行检测,按相同批次的试剂盒计算其批内CV;按相同批次的室内质控品不同批次的试剂盒计算其批间CV。结果乙型肝炎病毒表面抗原(HBsAg)酶联免疫诊断试剂盒和丙型肝炎病毒抗体(抗-HCV)酶联免疫诊断试剂盒,每种试剂均采用进口试剂与国产试剂同时进行检测。国产试剂的批内变异为:8.1%~17%,批间变异为:6.6%~19%;进口试剂的批内变异为:5.7%~12.2%,批间变异为:6.5%~8.3%。结论所使用的国产与进口试剂的批内和批间差异均控制在20%以内,能满足《血站技术操作规程》的要求。     

HBV-DNA实时荧光定量PCR检测系统性能验证

目的对国产PCR扩增仪、国产试剂实时荧光定量PCR检测HBV-DNA的性能作验证评价。方法以两质控血清按《医院检验科建设管理规范》方案作批内、批间不精密度评价;以定值标准血清重复3次测量,以偏差作准确性评价;以定值质控血清10倍倍比稀释,所得各样本重复3次测定,以示值与测定平均值作相关分析,作为线性评价;以定值标准血清样本,10次重复检测,以均能检测出阳性值(非阴性)为标准,评价最低检测限。结果两质控血清批内、批间精密度均5%;定值标准血清测量值对数值与认定值偏差不超过±0.5;线性评价相关性r=0.990,P0.000;最低检测限符合试剂盒说明。结论本实验室HBV-DNA荧光定量PCR检测系统性能符合YY/T 1182—2010《核酸扩增检测用试剂(盒)》要求,符合所用试剂盒说明的要求。     

总前列腺特异性抗原定量测定试剂盒(化学发光免疫分析法)的性能验证

目的对自制的总前列腺特异性抗原(total prostate specific antigen,t PSA)定量测定试剂盒(化学发光免疫分析法)进行性能验证,判断自制试剂盒是否满足产品行业标准要求。方法参照产品行业标准要求,对自制试剂盒的准确度、最低检测限、线性、重复性、批间差、稳定性、配对抗体等克分子反应性进行考察。结果自制试剂盒的最低检测限为0.0022 ng/m L,线性相关系数r=0.9998,重复性为2.82%和1.96%,批间差为5.55%,配对抗体等克分子反应小于15%,均符合产品标准的要求。对刚过12个月有效期的产品进行以上项目检测,也符合产品标准要求,说明效期内试剂质量稳定。结论自制试剂盒的各项性能指标符合行业标准要求,在线性、最低检测限和检测范围方面比临床常用的罗氏t PSA电化学发光试剂盒更优,可替代国外昂贵试剂应用于临床检测。     

HPLC法测定他克莫司滴丸的含量

目的建立他克莫司滴丸的含量测定方法。方法采用高效液相色谱法,色谱柱为Hypersil ODS(4.0mm×250mm,5μm),流动相为乙腈-水(60:40),柱温为50℃,流速1.0mL.min-1,检测波长为220nm;对3批他克莫司滴丸进行含量测定。结果他克莫司浓度在100～800μg.mL-1范围内,峰面积与浓度线性关系良好,平均回收率分别为99.92%(n=9),RSD为0.69%,3批样品中他克莫司的含量分别为99.76%、99.79%、99.93%。结论本法测定他克莫司的含量,简便快捷、结果准确、重现性好、易于操作,可用于他克莫司滴丸中他克莫司含量的测定。     

对31种乙型肝炎病毒表面抗原酶联免疫试剂盒的分析灵敏度评价

目的 评价乙型肝炎病毒表面抗原(HBsAg)酶联免疫(EIA)试剂盒的分析灵敏度.方法 对2007年6月至2008年6月申报批批检验的31种HBsAg EIA试剂盒,应用国家参考品进行检测,并用国家参考品中的灵敏度标准品,建立浓度和吸光度(A)值双对数曲线,将各试剂盒的Cut-off值代入曲线方程,计算其分析灵敏度,然后进行比较.结果 对27个国内厂家(351批)和4个国外厂家(27批)HBsAg EIA试剂盒进行分析,其中2个批次国产试剂盒的灵敏度低于国家要求的质量标准,总符合率为99.43%(349/351).国产试剂盒对adr、adw、ay血清型的分析灵敏度均值分别为0.307、0.419、0.513 ng/ml,差异有统计学意义(F=97.30,P<0.01);进口试剂盒的分析灵敏度均值分别为0.054、0.066、0.050 ng/ml,差异无统计学意义(F=0.65,P>0.05).进口试剂盒各血清型分析灵敏度均高于国产试剂盒(P<0.01).部分国产试剂盒与进口试剂盒对相同血清型的分析灵敏度差异有统计学意义.同一厂家试剂盒加酶后孵育30min和60min,其血清型分析灵敏度差异无统计学意义(P>0.05);同一厂家生产的试剂盒显色10 min和15 min,其血清型分析灵敏度的差异有统计学意义(P<0.01).结论 国产试剂盒应提高分析灵敏度,特别是对adw和ay血清型.     

他克莫司软膏治疗特应性皮炎的经济性评价

目的比较国产与原研药物他克莫司软膏治疗特应性皮炎的经济性,为临床选用治疗特应性皮炎外用药提供参考依据。方法通过药物释放与经皮渗透性研究对两种药品疗效和安全性进行比较;采用最小成本法对两种药品进行经济性比较。结果国产仿制药与原研药两种药品在疗效及安全性方面均无统计学差异;0.03%和0.1%他克莫司软膏国产仿制药比原研药费用分别减少31.29%和27.87%。结论他克莫司软膏国产仿制药治疗特应性皮炎与原研药相比,具有更好的经济性。     

酶联免疫反应加速仪在巨细胞病毒IgG抗体检测中的应用

目的:探讨酶联免疫反应加速仪在巨细胞病毒(CMV)IgG检测中的临床应用价值。方法:取标准品、阴性和阳性对照血清、已知结果的临床血清标本,使用酶联免疫收附试验(传统法)和酶联免疫反应加速仪法(加速法)进行巨细胞IgG检测,比较观察加速法检测CMVIgG的重复性、灵敏度和特异度是否符合试剂盒标准。结果:线性相关性分析加速法和传统法定量结果呈正相关关系。加速法的变异系数为10%～2.1%,灵敏度为98.2%,特异度为96.3%,均在规定范围内。结论:加速仪用于CMVIgG检测,各项指标均符合试剂盒要求,可在临床检测中使用。     

地奈德乳膏与他克莫司软膏序贯疗法治疗面部复发性皮炎疗效观察

目的 观察外用0.05%地奈德乳膏与0.1%他克莫司软膏序贯疗法治疗30例面部复发性皮炎临床疗效和安全性.方法 符合入选条件的60例面部复发性皮炎患者随机分为2组,各30例.实验组:先予外用0.05%地奈德乳膏早1次,0.1%他克莫司软膏晚1次,5d后改为每日早晚各擦1次0.1%他克莫司软膏,共10d.对照组:每日外用0.1%他克莫司软膏早晚各1次,连用10天.于治疗前及治疗后5d、10d观察临床疗效.结果 试验组疗效明显优于对照组,不良反应少.结论 应用0.05%地奈德乳膏与0.1%他克莫司软膏序贯疗法治疗面部复发性皮炎,安全、迅速、有效.     

自体表皮移植联合他克莫司软膏治疗稳定期白癜风疗效观察

目的观察自体表皮移植联合他克莫司软膏治疗稳定期白癜风的疗效及不良反应。方法将60例稳定期白癜风患者随机分成两组。治疗组:给予自体表皮移植联合他克莫司软膏治疗;对照组:单独给予自体表皮移植治疗。疗程均为3个月,疗程结束后根据照片进行临床疗效观察。结果治疗组有效率为93.33%,对照组为70%,两组间疗效比较差异具有统计学意义(P<0.05)。结论自体表皮移植治疗稳定期白癜风安全有效,联合外用他克莫司软膏能进一步提高疗效。     

戊型肝炎病毒IgG抗体酶联免疫诊断试剂盒研制

目的 研制戊型肝炎病毒IgG抗体（抗－HEV IgG）酶联免疫试剂盒（ELISA）。方法 应用2个人工合成的HEV多肽抗原包被微量反应板,用兔抗人IgG(γ链）辣根过氧化酶结合物作为第二抗体,检测血清中抗－HEV IgG,建立抗－HEV IgG ELISA;对该试剂盒的灵敏度、特异度、精密度、稳定性进行测定和临床考核。结果 经中国药品生物制品检定所检定,本试剂盒的灵敏度为90％（1／10）,特异度为100％（0／30）, 精密度＜15％,符合国家对抗HEV IgG ELISA的要求;在4℃下的稳定性至少为1年;与美国Genelabs公司和新加坡DBL抗－HEV IgG,ELISA总符合率分别为100％（43／43）和96．6％（86/89)。该试剂盒于1998年通过国家药品监督管理局的新药评审,获国家新药证书。结论 抗HEV IgG ELISA可用于戊型肝炎病毒感染的流行病学调查和临床诊断。     

鳞状细胞癌抗原单抗与免放试剂盒的制备及其临床价值

目的 制备鳞状细胞癌抗原(SCCA)单克隆抗体及免放分析试剂盒,并探讨其在宫颈癌、肺鳞癌临床诊断中的应用价值。方法 用重组人SCCA抗原免疫小鼠,通过细胞融合杂交瘤技术制备并筛选一对SCCA单抗,研制SCCA免放试剂盒,并评价其检测灵敏度,通过检测临床血清标本来评价其敏感度和特异性。结果 筛选出两株可以配对的SCCA单抗,研制的SCCA免放试剂盒检测灵敏度为0.08 ng/mL,对宫颈癌和肺鳞癌的临床测定的敏感度分布可达92.24%和89.52%,特异性为98.75%。结论 本SCCA免放试剂盒灵敏度高,特异性好,可用于宫颈癌、肺鳞癌的早期临床诊断和疗效观察,可有效提高宫颈癌、肺鳞癌的早期检出率。     

Evaluation of a panel of commercial kits for the detection of serum HIV-specific antibodies, and choice of alternative strategies for the serological diagnosis in Libreville (Gabon)

Nine commercially available kits for the screening of serum antibodies to the human immunodeficiency virus (HIV) were evaluated with a panel of 170 serum samples from adults in Gabon. The reference procedures showed that 96 samples had no antibodies, while 74 produced antibodies to HIV-1 (n=72) or HIV-2 (n=2). The sensitivity for 2 kits was less than 99% and the specificity of 3 less than 95%. Based on the panel of Gabonese serum samples, 4 of the 9 kits met the World Health Organization (WHO) acceptability criteria: sensitivity greater than 99% and specificity greater than 95%. Three kits available in Gabon that met these criteria were finally retained: Determine HIV-1/2, Genscreen Plus HIV Ag-Ab, and Immunocomb II HIV1&2 BiSpot. Of 18 configurations of alternative diagnosis strategies for HIV infection with these three kits, some WHO strategy II configurations (2 sequential assays) and all the WHO strategy III configurations (3 sequential assays) provided the maximum accuracy in diagnosing HIV infection in Gabon (sensitivity, specificity, positive predictive value, negative predictive value of 100%). The configuration using the Determine HIV-1/2 kit as first screening assay, followed by Genscreen Plus HIV Ag-Ab as a confirmatory assay, and finally, Immunocomb II HIV1&2 BiSpot as the third assay to discriminate samples discordant with the two first assays, was best, with high accuracy and less expense. The configuration of the two sequential rapid assays, Immunocomb II HIV1&2 BiSpot followed by Determine HIV-1/2, was also very accurate and reliable, as well as easiest to carry out (no need for ELISA technology), but it was twice as expensive as the first configuration with three sequential kits. In conclusion, when the laboratory facilities are available, the sequential diagnostic strategy of two rapid assays and a combined ELISA assay constitutes the best configuration, in terms of both accuracy and cost. When laboratory facilities are unavailable, sequential use of two rapid assays constitutes a convenient and accurate configuration, but is much more expensive.     

快速精子浓度检测试剂盒的临床实验室检测评价

目的:探讨快速精子浓度检测试剂盒的准确性、灵敏性和特异性,以评价其临床应用价值。方法:应用快速精子浓度检测试剂盒和世界卫生组织推荐的显微镜计数法分别测定临床500例不育症患者的精子浓度,并进行两种方法的kappa一致性检验。结果:快速检测精子浓度检测试剂盒和显微镜计数法比较的准确性为97.6%,特异性为97.4%,敏感性为97.8%,Kappa值为0.956,两种检测方法的结果有很好的一致性。结论:运用快速精子浓度检测试剂盒能够基本满足临床需求,其标本处理简便,有良好的应用前景。     

液相芯片技术检测卵泡液中细胞因子水平的可行性研究

目的:初步评价液相芯片技术用于检测控制性超促排卵患者取卵时卵泡液中36种细胞因子水平的可行性。方法:采用液相芯片技术检测81名不孕症患者单一卵泡卵泡液中36种细胞因子水平,通过检测结果初步评价其检测卵泡液中细胞因子水平的可行性。结果:液相芯片技术应用于检测卵泡液中白介素(IL)-1β、IL-4、IL-5、IL-10、瘦素(Leptin)、巨噬细胞炎性蛋白(MIP)-1β、肿瘤坏死因子(TNF)-β、血管内皮生长因子(VEGF)水平时,检测的准确度、精密度、敏感性均较高,结果可信。结论:液相芯片技术可用于检测卵泡液中细胞因子水平,但仍须进一步完善检测系统的评价及提高检验灵敏度,使可检测的卵泡液中目的细胞因子范围扩大,以提高其在卵泡液细胞因子水平检测中的应用价值。     

尿HB检测试剂盒在人群肿瘤筛检中的应用

为了探讨一种新的尿肿瘤标志物ＨＢ检测试剂盒在人群肿瘤筛检及肿瘤辅助诊断中的应用价值，对广州市各种原发性恶性肿瘤病人尿样进行双对照研究。结果表明，该试剂盒对１０多种常见恶性肿瘤检测总阳性率为６９０１％，明显高于良性疾病对照组（１０１４％）和健康人群对照组（４６２％，Ｐ＜０００１），但不同肿瘤类型尿ＨＢ阳性率似乎不同。检测灵敏度和特异度分别为８３０５％和９４９２％，诊断准确度为８８９８％。提示该试剂盒对常见恶性肿瘤的检测有较高的灵敏度和特异性，良性疾病因素对检测结果的干扰较小，并具有广谱、简捷、采样无创伤性等优点，可作为癌症的检测指标，在高危人群或职业人群肿瘤的筛检方面，以及作为恶性肿瘤的辅助诊断方面均有较好的应用价值     

Rapid spectrophotometric determination of plasma carnitine concentrations

A spectrophotometric enzymatic assay for plasma carnitine concentrations has been automated on the Monarch 2000. Prior to the assay, each plasma sample was divided into three fractions, ie, free carnitine, acid-soluble carnitine and total carnitine, in order to determine the concentration of both free and esterified carnitine. Using the method developed by Tachikawa et al (Seikagaku 56:998, 1984), each of the samples was then chromatographed on an anion exchange resin to eliminate those compounds which could adversely impact the accuracy of the enzymatic assay for carnitine. After the completion of these preparatory steps, 32 specimens were assayed in less than 16 min on the Monarch 2000 with a high degree of both accuracy and precision. The assay was linear over a wide concentration range (5.0-80 mumol/liter), with the lower limit of sensitivity being 5.0 mumol/liter. The coefficient of variation (CV%) of the within run precision was 2.1%, 2.8%, and 6.7% for the determinations of free carnitine, acid-soluble carnitine, and total carnitine, and 17.4% and 27.8% for the calculated values of short-chain and long-chain acylcarnitines, respectively. The CV% of the between run precision for the same fractions was 6.5%, 2.7%, 3.8%, 14.2%, and 14.3%, respectively. When authentic L-carnitine was added to the plasma, the mean recovery rate was 94.7 +/- 11.0%. Reference values were determined using plasma obtained from 40 healthy adult volunteers.     

快速精子浓度自检试剂盒性能评价

目的:评价快速精子浓度自检试剂盒的性能。方法:对7个批号的快速精子浓度自检试剂盒进行灵敏性、特异性、准确性、稳定性等指标检测。结果:7批试剂盒灵敏性为96.7%,特异性为96.1%,准确性为96.3%,稳定性>12个月。结论:快速精子浓度自检试剂盒灵敏性、特异性、稳定性均达到要求,且具有良好的准确性和重复性。     

Development and field validation of an avidin-biotin enzyme-linked immunosorbent assay kit for bovine brucellosis.

The avidin-biotin enzyme-linked immunosorbent assay (A-B ELISA), for use in surveillance for bovine brucellosis in India was developed and calibrated using the indirect brucellosis ELISA kit of the International Atomic Energy Agency (IAEA) as a reference. The reagents used in the A-B ELISA were as follows: the smooth lipopolysaccharide of Brucella abortus strain 99 (antigen); biotinylated anti-bovine immunoglobulin G (detection antibody); avidin-horseradish peroxidase (conjugate); and O-phenylenediamine dihydrochloride (chromogen). The test results were interpreted using the IAEA software EDI version 2.1.1, which was modified for use in the A-B ELISA. The cut-off percentage positivity value was established using 500 brucellosis-positive and 500 brucellosis-negative serum samples, confirmed with reference to the sample data using the indirect ELISA kit. The overall specificity of A-B ELISA was 98.8% and overall sensitivity was 98.2%. Field validation of the A-B ELISA kit was undertaken in six laboratories in India. Screening of 7,040 cattle and 678 buffalo serum samples from 12 states revealed serological evidence of brucellosis in 8.7% of cattle and 10.2% of buffalo. This kit proved to be robust and performed with a similar sensitivity and specificity to the indirect ELISA. The kit can be supplied at a lower cost than current commercial ELISA kits.