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1.
新型戊型肝炎诊断试剂盒的研制及其应用   总被引:5,自引:1,他引:4  
用HEVORF3合成肽及ORF2重组抗原研制成新型HEVEIA诊断试剂盒。与GenlabsHEVEIA检测比较,灵敏度和特异性均达100%(60/60)。三批试剂精密性测定均<10%。该试剂盒置4℃8个月或37℃4d保持稳定。检测不同肝炎患者HEV抗体,发现急性非甲非乙非丙肝炎中有63.2%,甲肝有13.4%,乙肝有8.3%,丙肝有6.6%,正常人群为2.9%。所研制的戊型肝炎诊断试剂盒,灵敏度高,特异性强,精密性好,稳定性合格。适用于戊型肝炎诊断及戊肝病毒感染的流行病学调查  相似文献   

2.
鼠颌下腺提纯的25SNGF免疫家兔,获得兔抗NGF抗体,研制出鼠25SNGFELISA检测试剂盒,该试剂盒灵敏度小于1ng/ml,在670~084ng/ml范围内,线性良好,r=099。与大鼠、小鼠及人血浆无非特异反应,在大鼠血浆中,NGF样品回收率在91%~107%之间,变异系数小于10%(n=4)。结果表明:本试剂盒操作简便,灵敏度高,特异性强,适合药代动力学研究及生产过程中的NGF检测。  相似文献   

3.
轮状病毒乳胶试剂盒的研制   总被引:1,自引:0,他引:1  
用轮状病毒A群单克隆抗体研制出轮状病毒抗原乳胶凝集试剂盒,并与轮状病毒ELISA试剂盒进行了比较。试验结果表明,轮状病毒抗原乳胶凝集试剂盒具有较高的特异性(987%)和敏感性(868%),与ELISA试剂盒比较无显著性差异(P>01),且操作简便快速、成本低廉  相似文献   

4.
用DIGFA单盲检测各类胃病患者血清HP特异性抗体,结果表明其敏感性为979%,特异性为915%,诊断效率达947%,youden’s指数为089。通过本法与常规ELISA比较,发现两法前四种诊断指标均无明显差异,但DIGFA简易、快速,无需任何设备,数分钟内即可得出正确结论,因此适合于基层医院查病、胃镜检查前过筛及流行病学调查应用  相似文献   

5.
用聚合酶链式反应(PCR)从HIV 1gag基因中扩增出衣壳蛋白P24截短体(tP24)基因,插入质粒pGEX 4T3中构成重组表达质粒pGEX tp24,将pGEX tp24转化大肠杆菌BL21后获得了高效表达,表达量占菌体总蛋白量的3438%。经Glutathione-Sepharose4B亲和层析纯化的截短体P24纯度为9277%。纯化的截短体P24在间接ELISA和免疫印迹检测HIV抗体阳性血清和正常人血清中,具有很高的抗原特异性和免疫反应性。  相似文献   

6.
陈卫  付必谦 《兽类学报》1994,14(4):312-313
中华姬鼠种群生态的初步分析THEPRELIMINARYANALYSISONTHEPOPULATIONECOLOGYOFCHINESEFIELDMOUSE(APODEMUSDRACO)KeyWordsChineseFieldMouse(Apodemus...  相似文献   

7.
报道了用辣根过氧化物酶标记的抗人IgG和抗人IgM(u链)单克隆抗体作第二抗体,用自己培养、纯化的弓形体(To)、风疹病毒(RuV)、巨细胞病毒(CMV)和单纯疱疹病毒(HSV12)的虫体和病毒抗原包被酶标板,研制出检测ToRCH系列的特异性IgG和IgM的间接ELISA试剂。质量检定结果表明,该试剂特异性强、本底低,能有效消除RF因子等干扰因素的影响;灵敏度达1∶160~640;精密性好,变异系数(C.V)在1.4%~9.0%;试剂稳定,37℃存放4d,各项指标的变化率不超过15%。  相似文献   

8.
本文报道对清洁级实验动物应排除的四种病毒(淋巴细胞脉络丛脑膜炎病毒、小鼠脱脚病病毒、鼠肝炎病毒和仙台病毒)抗体玻片酶免疫(EIA)检测试剂盒的研制。四种病毒感染的细胞和对照细胞经冷丙酮固定于载玻片上制成特异性抗原和对照抗原,此四种病毒的抗血清各10份和SPF小鼠血清20份分别与四种病毒的特异性抗原和对照抗原进行EIA交叉试验,结果显示,抗原只与其相应抗血清发生特异性显色反应,与非特异性小鼠血清和SPF小鼠血清不显色。与HI或ELISA方法比较,通过对112份普通小鼠血清进行测试,结果表明,EIA对仙台病毒抗体的检出率(19.6%)显著高于(<0.005)HI(6.3%),对小鼠脱脚病病毒抗体的检出率(23.3%)与HI(21.4%)无显著性差异(P>0.05)。EIA对淋巴细胞脉络丛脑膜炎病毒和鼠肝炎病毒抗体的检出率分别为1.8%和71.2%,ELISA对两种病毒抗体的检出率分别为1.8%和67.6%,两种方法对两种病毒抗体的检出率无显著性差异(P>0.05)。重复性试验表明两批四种病毒抗体试剂盒对108份小鼠血清两次测定的符合率为96~100%。四种病毒的EIA抗原在-18℃保存12个月或在2-8℃保存3  相似文献   

9.
拟南芥:植物分子生物学研究的模式物种   总被引:7,自引:0,他引:7  
陈璋 《植物学通报》1994,11(1):6-11
拟南芥:植物分子生物学研究的模式物种陈璋(福建省花卉研究中心福州350002)ARABIDOPSISTHALIANAASAMODELSPECIESFORPLANTMOLECULARBIOLOGYSTUDIES¥ChenZhang(FujianProu...  相似文献   

10.
乙型肝炎诊断试剂国家质控标准的建立   总被引:1,自引:0,他引:1  
按新的方案建立了用于乙型肝炎诊断试剂的国家质控标准。包括用WHO国际标准品标化建立的我国HBsAg、抗-HBs定量标准品,对试剂的特异性、灵敏度、精密性、线性进行质量控制检定的系列参比品,及经过实验制订的检定标准。经近年来的实际应用,对国产试剂质量的提高起到了推动作用。  相似文献   

11.
Laboratory scale development of a two site micro enzyme linked immuno assay kit is described. The kit comprises rabbit anti human alphafetoprotein (AFP), anti human AFP IgG peroxidase conjugate and standard AFP. All the above reagents were prepared in the laboratory. The kit is eminently suitable for early screening of blood sample of pregnant women for neural tube defects of their fetuses and for the quantitation of AFP as a tumor marker. The assay kit was used to determine AFP in 76 sera from women at different stages of pregnancy. During 1st trimester AFP level was 18 to 119 ng/ml, during 2nd trimester the concentration varied from 85 to 302 ng/ml and during 3rd from 103 to 580 ng/ml. No evidence for maternal antibody to AFP was found. The above data agree with AFP level in pregnant women reported by earlier workers, using RIA or ELISA. The present ELISA kit would hopefully be much cheaper than internationally available ELISA kits for human AFP.  相似文献   

12.
摘 要:目的 对本实验室乙型肝炎病毒核酸(HBV DNA)定量检测程序方法学性能进行评价,以了解其方法学性能是否符合临床及本实验室检测要求。方法 分别取高浓度(HP)和低浓度(LP)两个水平的患者标本对精密度进行评价,取国家标准品(L1-L6)对正确度、线性范围及检测下限进行评价。结果 高浓度(HP)和低浓度(LP)两个水平的患者标本CV均低于5.0%,精密度符合要求;L1-L6检测结果均在国家标准品的允许范围内,正确度符合要求;a=0.9867,r2=0.9985,线性范围符合要求;国家标准品中的L4稀释后,300 IU/mL、500 IU/mL均可检出,符合试剂盒的检测灵敏度要求。结论 本室所评价的试剂盒方法学性能指标均符合要求,适合在本室开展。  相似文献   

13.
目的评价国产小鼠病毒抗体ELISA检测试剂盒。方法选择国产与进口小鼠淋巴细胞脉络丛脑膜炎病毒(LCMV)、肝炎病毒(MHV)、仙台病毒(SV)、腺病毒(MAV)、细小病毒(MPV)ELISA抗体检测试剂盒,进行敏感性、特异性、精密性、稳定性、可信度试验比较。结果国产与进口试剂盒:同种试剂盒之间灵敏度相差最低为2倍,差异显著(P〈0.05),最高为16倍,差异极显著(P〈0.01);特异性试验显示每种试剂盒,与其他4种病毒均无交叉反应;精密性试验显示5种试剂盒批内平均变异系数均小于10%;稳定性试验显示5种试剂盒相对偏差均小于25%;分别选择已知36份小鼠血清进行检测,国产和进口LCMV、MHV、SV、MPV符合率均为100%;国产MAV符合率为86.1%,进口MAV符合率均为100%,二者之间差异极显著(P〈0.01)。结论除国产MAV试剂盒敏感性、可信度低于进口外,国产LCMV、MHV、SV、MPV试剂盒与进口同种试剂盒相比,在敏感性、特异性、精密性、稳定性和可信度方面均良好。  相似文献   

14.
The results of comparative study of sera obtained from donors and from several groups of patients with suspected toxoplasmosis are presented. The study has been carried out with the use of commercial enzyme immunoassay (EIA) kits: Toxoplasma gondii IgG EIA and Toxoplasma gondii IgM EIA manufactured by Labsystems (Finland), Sevatest ELISA IgG Toxo Micro I manufactured by Sevac (Czechoslovakia). Statistical processing of the results has confirmed the identity of these kits. The necessity of using evaluation criteria (the separation point, the scale for the interpretation of results) when working with the Sevac kits is emphasized. Comparative evaluation of antibody profiles in the sera under test suggests that the titer less than 1:1600 should be regarded as the separation point for these kits. IgM antibodies to T. gondii have been found only in 22% of patients with high titers of IgG antibodies.  相似文献   

15.
At the Institute of Sera and Vaccines, Praha, was invented and tested on clinical samples a kit for detection and quantification of alpha 1 fetoprotein in human serum. It is a heterogeneous EIA on the "sandwich" principle. Rabbit antibody to alpha 1 fetoprotein (further AFP) was used for coating the solid surface and goat horse-radish peroxidase labelled antibody to AFP was used as the tracer. Microtitration plate of Czechoslovak manufacture (KOH-I-NOOR, Dalecín) type P with 96 wells was used as the solid phase. The range of an approximately linear part of the calibration curve was intentionally chosen between 10 and 400 ng/ml, since in this way it fills the detection gap in AFP determination between 10 and 200 ng/ml, which is, on the one hand, a physiological value of AFP in human serum and, on the other hand, the bottom limit of sensitivity of counter immunoelectrophoresis (CIEP). Attention was devoted both to reproducibility of the method, i.e. results of intra- and interassays, and comparability with other foreign ELISA Kits. According to the correlation analysis, the kit was ascertained to be very well comparable with kits of foreign provenance. The coefficient of variation (CV) for the interassays varied between 11 and 16% and for intraassays it equalled 15%.  相似文献   

16.
An analytical procedure to screen butorphanol in horse race urine using ELISA kits and its confirmation by GC–MS is described. Urine samples (5 ml) were subjected to enzymatic hydrolysis and extracted by solid-phase extraction. The residues were then evaporated, derivatized and injected into the GC–MS system. The ELISA test (20 μl of sample) was able to detect butorphanol up to 104 h after the intramuscular administration of 8 mg of Torbugesic, and the GC–MS method detected the drug up to 24 h in FULL SCAN or 31 h in the SIM mode. Validation of the GC–MS method in the SIM mode using nalbuphine as internal standard included linearity studies (10–250 ng/ml), recovery (±100%), intra-assay (4.1–14.9%) and inter-assay (9.3–45.1%) precision, stability (10 days), limit of detection (10 ng/ml) and limit of quantitation (20 ng/ml).  相似文献   

17.
By using an improved hybridoma technique, monoclonal antibodies specific to alpha-fetoprotein (AFP) were generated. After three subcutaneous immunizations and three intravenous boosters, cell fusion experiments were performed. The hybrid cells were initially cultured in a semisolid medium containing methylcellulose and later transferred to a liquid medium for subculture. Out of 800 colonies recovered after two cell fusion experiments, 16 were shown to exhibit affinity to AFP by radioimmunoassay. Six hybrid cell lines which showed high affinity and specificity were selected for further evaluation. From the results of a cross-matching procedure, two pairs of antibodies (AFP 3 and AFP 05; AFP 3 and AFP 013) reacting with discrete antigenic determinants were identified for preparing solid-phase sandwich enzyme immunoassay (EIA) kits. The association constants between AFP and these three antibodies (AFP 3, AFP 05, and AFP 013) were 2.0, 3.7, and 3.8 X 10(9) M-1, respectively. The immunoglobulin subclass of them was determined to be IgG1. The EIA procedure designed could be performed within 40 min in a one-stage incubation and 70 min in a two-stage incubation. The incubation time was shown to be equal to or shorter than that of any other known commercial kits and the sensitivity was less than 1 IU/ml. In order to avoid the high-dose hook effect which occurred in the one-stage incubation procedure, a two-stage incubation protocol was advised.  相似文献   

18.
Alpha-fetoprotein (AFP) is a diagnostic marker for hepatocellular carcinoma (HCC). A direct relationship between poor prognosis and the concentration of serum AFP has been observed. Telomerase, an enzyme that stabilizes the telomere length, is expressed by 90% of HCC. The aim of this study was to investigate the effect of telomerase inhibition on AFP secretion and the involvement of the PI3K/Akt/mTOR signaling pathway. Proliferation and viability tests were performed using tetrazolium salt. Apoptosis was determined through the Annexin V assay using flow cytometry. The concentrations of AFP were measured using ELISA kits. The AFP mRNA expression was evaluated using RT-PCR, and cell migration was evaluated using a Boyden chamber assay. The in vivo effect of costunolide on AFP production was tested in NSG mice. Telomerase inhibition by costunolide and BIBR 1532 at 5 and 10 μM decreased AFP mRNA expression and protein secretion by HepG2/C3A cells. The same pattern was obtained with cells treated with hTERT siRNA. This treatment exhibited no apoptotic effect. The AFP mRNA expression and protein secretion by PLC/PRF/5 was decreased after treatment with BIBR1532 at 10 μM. In contrast, no effect was obtained for PLC/PRF/5 cells treated with costunolide at 5 or 10 μM. Inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP concentration. In contrast, the MAPK/ERK pathway appeared to not be involved in HepG2/C3A cells, whereas ERK inhibition decreased the AFP concentration in PLC/PRF/5 cells. Modulation of the AFP concentration was also obtained after the inhibition or activation of PKC. Costunolide (30 mg/kg) significantly decreased the AFP serum concentration of NSG mice bearing HepG2/C3A cells. Both the inhibition of telomerase and the inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP production of HepG2/C3A and PLC/PRF/5 cells, suggesting a relationship between telomerase and AFP expression through the PI3K/Akt/mTOR pathway  相似文献   

19.
The prevalence of chronic Toxoplasma infections reported in the literature varies enormously. We hypothesize that one factor could be due to the different methods used in the evaluation of infections. Serological evidence of Toxoplasma infections in 450 pregnant women (PW) and 300 HIV-infected patients (HIV) were investigated by the Sabin–Feldman dye test and two other commercial ELISA kits (kit1 and kit2). Anti-Toxoplasma IgG antibodies obtained from the Sabin–Feldman dye test, ELISA kit1 and ELISA kit2 in the PW subjects were 14.7%, 29.6% and 38.7%, and in the HIV subjects were 13%, 34.7% and 36.3%, respectively. So there were significant differences in the seroprevalences when different diagnostic tests were used (P < 0.05). Regarding Sabin–Feldman dye test as the gold standard for anti-Toxoplasma antibodies detection, we found that the sensitivity and specificity of the ELISA kit1 and kit2 was in the range of their specification. However as the two ELISA kits used in our study identified a much higher prevalence of Toxoplasma infections which indicated that false positive cases were being reported. Based on results obtained, it is therefore highly recommended that research workers should be aware that the reports of serological studies in terms of high positive results should be treated with some skepticism until additional precise diagnostic tools are developed.  相似文献   

20.
Measurement of chromogranin A (CgA) plays a major role in the management of neuroendocrine tumors (NET); however, reliable assaying of CgA is made difficult by the rapid hydrolysis following its release into the bloodstream. This study was aimed at the assessment of two assays for CgA in NET patients. CgA was measured in 93 patients by means of an enzyme-linked immunosorbent assay (ELISA) and an immunoradiometric assay (IRMA). The specificity and sensitivity of CgA were evaluated in relation to tumor histology. The clinical accuracy of the two assays was evaluated by receiver-operating characteristic (ROC) curve analysis. Regression analysis demonstrated different immunoreactivity for CgA of the antibodies used in the two kits (r = 0.61). The two assays had different accuracy also in classifying patients according to their clinical condition (91% vs 64% specificity and 79% vs 79% sensitivity for the ELISA and IRMA assay, respectively) and tumor histology (81% vs 85% sensitivity for the ELISA and IRMA assays, respectively, in carcinoids; 92% vs 67% sensitivity for the ELISA and IRMA assays, respectively, in pancreatic islet cell tumors). The different clinical accuracy of the two assays was confirmed by the ROC analysis (AUC = 0.90 vs AUC = 0.87 for the ELISA and IRMA assays, respectively). In conclusion, because of the poor standardization of the commercially available measurement tools the clinical accuracy of CgA measurement depends on the assay used. This makes it difficult to compare CgA values measured with different kits and affects the clinical accuracy of the different assays for CgA.  相似文献   

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