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1.
用兔抗人IgG4的多克隆抗体(IgG4Ab)包被成固相板,以识别结合待测标本中的IgG4。用生物素标记IgG4Ab得BioIgG4Ab;用亲和素标记辣根酶得HRPA。在已包被IgG4Ab的固相板中加入IgG4标准品(或待测样品),反应、洗涤后,加入BioIgG4Ab,反应、洗涤后,加入HRPA,反应、洗涤后,板上形成IgG4Ab—IgG4—BioIgG4Ab—HRPA复合物,加酶底物显色,用酶标仪在490nm波长测定OD值,作标准曲线,根据标准曲线,查出标本中IgG4含量。该法测定范围:2.5~200ng/mL,最低检出量2.1ng/mL,批内和批间CV分别8.4%和11.2%。测得血清IgG4含量:青年人(30名)为37.7±2.3ng/mL;30名献血员为42.7±9.9ng/mL,49例重症监护患者明显升高为71.2±9.2ng/mL;49例肺部感染患者明显降低为28.4±9.2ng/mL。结果表明该法稳定,灵敏度适于检出人血清IgG4水平。  相似文献   

2.
目的:建立乳铁蛋白(LF)酶免疫分析法,检测LF在血清及乳汁中的水平。方法:用人源的LF多次免疫兔,获得高效价的抗体。纯化后包被成固相酶标板,以识别并结合待测标本中的LF。用生物素标记抗原(Bio-Ag),同辣根酶标记亲和素(HRP-A)可特异结合。向固相酶标板中同时加入Bio-Ag和不同浓度的标准品,37℃反应1.5h。洗涤后以HRP-A为探针,经酶底物显色后可获得良好的ELISA竞争抑制曲线。结果:该法测定范围:(0.36~9.00)μg/ml,最低检出量0.25μg/ml,批内和批间误差分别为〈6.3%和7.4%。用该法测正常献血员(40名)血清LF水平为(0.438±0.183)μg/ml;人初乳(5名)水平为(7.82±0.51)mg/ml。血清和初乳中的LF经65℃30min处理后活性不变,经100℃3min处理明显降解。结论:该法稳定、简便、特异适于检测人血清和乳汁中的LF水平。  相似文献   

3.
人血清抵抗素ELISA方法的研究及其临床初步应用   总被引:1,自引:0,他引:1  
用兔抗人抵抗素(resistin)片断(22~34氨基酸残基)的多克隆抗体(PcAb)包被固相板,用辣根过氧化物酶(HRP)标记PcAb得HRP-PcAb.在包被PcAb的孔中加入抵抗素标准品(或待测样品),加入HRP-PcAb,形成PcAb-resistin-HRP-PcAb复合物,加酶底物邻苯二胺(OPD)显色,用酶标仪在492nm波长处测定OD值,绘制标准曲线.从标准曲线读出样本中抵抗素含量.本方法特异性强,灵敏度高,精密度好.该法测定范围0.5~100ng/mL,最低检出量0.5ng/mL,批内和批间CV%分别为3.4%和7.2%.50例2型糖尿病患者血清抵抗素含量为23.2±3.9 ng/mL,明显高于正常值16.0±2.5ng/mL.结果表明该法稳定,灵敏度适于检测人血清抵抗素水平.  相似文献   

4.
目的:探讨血清对氧磷酶-3(PON3)和载脂蛋白A-I(apoA-I)糖氧化水平与2型糖尿病(T2DM)并发冠心病(CAD)及其严重程度的关系。方法:202例T2DM患者按其是否合并CAD分为单纯糖尿病组(DM组,n=60)和T2DM合并CAD组(DM并CAD组,n=142),后者进一步按病变累及冠脉支数分为1支病变组(n=40),2支病变组(n=52)和3支病变组(n=50)。另选62例健康受试者为对照组。采用ELISA测定各组血清PON3水平;超速离心全血获得高密度脂蛋白(HDL),采用SDS-PAGE电泳分离apoA-I,免疫印迹分析apoA-I糖氧化水平。分析PON3水平和apoA-I糖氧化水平与冠脉病变程度的相关性。结果:DM并CAD组和3支病变组血清PON3水平(1.52±1.14ng/ml、1.31±1.09ng/ml)分别显著低于DM组(1.98±1.04ng/ml,P0.05)、对照组(2.46±1.01ng/ml,P0.05)和1支病变组(1.74±1.19ng/ml,P0.05)。apoA-I糖氧化水平是CAD及其病变严重程度的独立预测因子,并与冠脉病变严重程度指数、病变冠脉支数呈显著正相关(r分别为0.611、0.549,P均0.01)PON3水平与apoA-I糖氧化水平呈负相关(r=-0.388,P0.01),且两者均为CAD独立预测因素。结论:低PON3水平与apoA-I糖氧化水平相关,后者与T2DM患者并发CAD及其严重程度相关。  相似文献   

5.
本测定所用方法为以聚苯乙烯反应板为载体的双抗体夹心法。用于包被反应板及酶标记的抗HBeIgG系用DEAE-纤维素层析法从抗HBe阳性血清中提取而得。检测的操作步骤如下:(1)用0.75%鞣酸溶液处理聚苯乙烯反应板。43℃,1小时。洗涤。(2)加入抗HBe IgG(20~25微克/毫升)100微升。4℃过夜。洗涤。(3)在测定孔中加入血清标本50微升,正常人血清50微升;阳性对照孔中用HBeAg阳性血清代替血清标本;阴性对照孔中只加正常人血清。43℃,1小时。洗涤。(4)加入用含  相似文献   

6.
白介素8酶联免疫分析方法及其初步应用   总被引:1,自引:0,他引:1  
建立高灵敏度和特异性的白介素8(IL-8)ELISA法.用人工重组的IL-8多次免疫兔,获取高效价抗体.纯化抗体经生物素标记,同辣根过氧化酶(HRP)标记的亲和素可特异结合.采用纯化的抗体包被96孔板,形成固相抗体.加入不同浓度标准品后,反应1.5h,洗净后再加入生物素标记抗体,以酶标记亲和素作为探针,经酶底物显色反应可获得良好的ELISA标准曲线.该方法测定范围为0.2-10.8ng/mL,最低测出值为0.02ng/mL,批内和批间CV小于8%和10%.用该法测得18名正常人血浆中IL-8水平为0.15±0.06ng/mL.18份高低不同浓度IL-8水平的血清样品经本法和RIA测定其均值为0.54±0.35ng/mL和0.63±0.59ng/mL,相关性强(r=0.819,t=5.89).在U937细胞系体外培养液中,LPS刺激24h和48h可测出IL-8水平明显上升;而经TNF-α刺激24h和48h后同对照没有明显变化.该方法操作简便,具有较强灵敏度和特异性,可满足人血清和细胞培养液中IL-8水平的检测.  相似文献   

7.
高滴度抗HBc IgG从HBsAg阳性(抗HBs阴性)肝炎患者血清提取。制备抗HBc·HRP(抗HBcIgG或根过氧化物酶结合物)按戊二醛二步法辣过碘酸钠改良法。试剂HBcAg从阳性尸肝提取。ELISA抑制法检测抗HBc时是将试剂HBcAg先结合在已用抗HBc IgG包被的聚苯乙稀板孔内,加入待检血清经反应和洗涤,再加入抗HBc·HBP经反应和洗涤,  相似文献   

8.
山西汉族可溶性HLA-Ⅰ类抗原检测   总被引:1,自引:0,他引:1  
目的:定量检测正常人血清中可溶性人白细胞抗原-Ⅰ(sHLA-Ⅰ),以探讨山西汉族人群的正常参考值.方法:ELISA法检测血清sHLA-Ⅰ类抗原水平.将不同浓度的标准品和待检血清分别加入包被有特异性sHLA-Ⅰ抗体的酶标板,与生物素化的sHLA-Ⅰ、辣根过氧化物酶(HRP)标记的链霉亲和素共同孵育,弃上清并用洗涤液洗涤,加底物应用液四甲基联苯胺(TMB)显色,以酶标仪450nm测定吸光度值,根据不同浓度标准品显色后测得的吸光度值绘制标准曲线,测定60例健康山西人的sHLA-Ⅰ含量.结果:ELISA法能够准确检测sHLA-Ⅰ含量,60例健康山西人的sHLA-Ⅰ含量为(382.62±106.68)ng·ml-1.结论:山西汉族人群sHLA-Ⅰ正常值为(382.62±106.68)ng·ml-1.  相似文献   

9.
作者建立了一种检测结核分枝杆菌抗原的双抗体夹心间接ELISA的方法并介绍了其应用情况。用鼠源性抗结核杆菌单克隆抗体包被酶标反应板,依次加入经0.5%胰蛋白酶消化的肺结核病人痰液标本,人免抗结核杆菌超免疫血清和酶标羊抗兔IgG孵育后(每步均经孵育,洗涤),加底物显色。试验中采用结核杆菌Ho7RV株聚合OT免疫家兔制备的超免疫血清作为桥联抗体,既为试验的特异性奠定了基础,也避免了标记IgG的繁冗操作,减少了IgG的活性损失。  相似文献   

10.
为使ELISA试验结果稳定、敏感,有报告在ELISA反应液中加入聚乙二醇(PEG)能加速病毒及细菌抗体的测定,并有助于灵敏度的提高。本文将此法试用于对乙型肝炎表面抗原(HBsAg)的测定,其方法简述如下。 一、抗体包被:在聚苯乙烯微量反应板的每孔内加入0.1毫升用包被缓冲液(pH9.6,0.05M碳酸钠-碳酸氢钠缓冲液)稀释至含量为10微克/毫升的羊抗HBs,置4℃冰箱过夜,用洗涤缓冲液(pH7.4,0.02MTris-Tween 20)洗涤三次。 二、样品反应:每孔加入用稀释缓冲液(含4%PEG的pH7.4 PBS-Tween 20)作1:10稀释的血清0.1毫升,每份样品同时测2孔,置43℃1小时,洗涤同上。 三、酶标记物反应:每孔内加入用稀释缓冲液适当稀释的过氧化物酶标记羊抗-HBs0.1毫升,置43℃1小时,洗涤同上。  相似文献   

11.
A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.  相似文献   

12.
背景:局部注射具有成骨作用的辛伐他汀,可显著增加骨质疏松大鼠股骨颈及股骨髁部的骨密度及力学强度,分析局部注射辛伐他汀对股骨髁骨小梁的影响。 目的:进一步研究骨质疏松大鼠股骨内局部注射辛伐他汀对股骨髁骨小梁的影响。为将辛伐他汀应用于临床骨质疏松局部治疗提供实验基础。 方法:18只雌性SD大鼠双侧卵巢切除后3个月,制备大鼠骨质疏松模型。实验大鼠随机数字表法均分为3组,分别在实验大鼠的右侧股骨髓腔内单次注射辛伐他汀溶液5 mg、10 mg,对照组单纯注射空白载体。分别在注射后1个月处死大鼠并取材。Micro-CT扫描并定量分析骨组织形态变化。 结果与结论:给药后1个月,Micro-CT扫描结果显示,辛伐他汀治疗组的骨微结构参数如骨皮质厚度、骨小梁密度及连接率明显优于对照组。说明疏松骨骼单次注射小剂量辛伐他汀可显著促进股骨髁部骨小梁改建,改善骨骼微结构,可为强化局部、防治骨质疏松骨折的新选择进一步提供实验基础。  相似文献   

13.
石斛属民族药用植物的分类及生药学研究   总被引:2,自引:0,他引:2  
中药石斛早在《神农本草经》中就被列为上品,其应用历史悠久,具有养阴生津、补肾益气、润喉护嗓、活血明目、抗癌防老等功效。本文对国内外石斛属民族药用植物的分类及生药学研究进行了综述。  相似文献   

14.
目的我科自2001~2005年4月对59例67膝的骨性关节炎(0A),进行关节镜检查及镜下清理术。方法镜检:膝关节骨性关节炎伴不同程度滑膜炎,滑膜皱壁粘连纤维片,关节软骨不同程度破坏。镜下清除增生滑膜,松解粘连,去除剥脱软骨、修复关节面。结果随访2月~4年,平均1.5年,术后综合评估2月~1年,优良率94%;1~2年,优良率79.2%;2年以上优良率54.8%。结论关节镜对膝关节骨性关节炎诊断能提供了比较全面的资料,并对骨性关节炎早期有良好疗效,具有创伤小、恢复快、并发症少和重复治疗等优点。  相似文献   

15.
Since the cutaneous veins of the four limbs have been used as autogenous grafts in the treatment of cardiovascular disorders, knowledge of distribution of the valves is increasingly required depending upon the use. In the gross anatomical study of distribution of valves of the trunci venae, there has been argument about locational relationships between the venous roots and the valves in the vicinity of the roots and the inter-valvular distance. However, there have been only few reports discussing detailed information about valves of the cutaneous veins of the four limbs. The authors observed patterns of distribution of the cutaneous venous valves of the four limbs of cadavers prepared for practice in anatomy. The following parts were excised from each cadaver: the cephalic, basilic, and the great saphenous veins, which originate from the acral venous network and flow into the proximal deep veins, and the venous roots communicating with these veins. An incision was made on each excised vein in the direction of the long axis under observation with a stereoscopic microscope, and the inter-valvular distance and the distance between the valve and the orifice of venous root in the vicinity of the valve were measured. The inter-valvular distance varied with type of the truncus venae, and it varied according to area even in the same truncus venae.  相似文献   

16.
Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 115, N o 4, pp. 401–404, April, 1993  相似文献   

17.
Sterols of three digenetic trematodes were isolated and characterized by infrared and 1H nuclear magnetic resonance spectroscopies, and gas-liquid chromatography and gas chromatography-mass spectrometry. Sterols identified were cholesterol, cholestanol, 24-methylcholesterol, 24-methylcholestanol, 24-ethyl-22-dehydrocholesterol, 24-ethyl-22-dehydrocholestanol, 24-ethylcholesterol and 24-ethylcholestanol.  相似文献   

18.
New electrophysiologic devices were used for the diagnostics of the state of excitable structures in the gastrointestinal tract and correction of their motor functions. Bioelectrical and biomechanical activities form the basis of functioning of internal organs. The mechanisms of generation of slow bioelectrical activity that are important for clinical and physiological studies are described. One of these mechanisms is a capacitance parametric transducer converting the energy of contractions into specific electric signals reflecting muscle functions. Another mechanism results from slow oscillations of resting potentials of interrelated excitable cells in large neuromuscular structures of internal organs. The elaborated procedure is efficient for preventing early postoperative paresis of the gastrointestinal tract. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 10, pp. 448–452, October, 1999  相似文献   

19.
崔慧  董鹏  刘文娟  徐敏 《解剖与临床》2008,13(4):239-240
目的:研究正常胃小弯侧血管弓及其属支血管的多排螺旋CT表现。方法:回顾性分析51例正常成人的上腹部CT资料,观察胃小弯侧血管弓及其属支血管的显示情况、血管弓形态及其与胃壁的关系。结果:51例均显示胃小弯侧血管弓,血管弓贲门部属支、胃体部属支和幽门部属支分别显示46例、15例和10例;血管弓与胃壁紧邻41例。结论:MDCT检查可良好显示正常胃小弯侧血管弓及其属支血管。  相似文献   

20.
Assessment of the aftereffects of cryoexposure and ultrahigh-frequency cryoexposure on hemangioma tissue of various types, cavernous and squamous, showed a higher cryogenic effect in hemangiomatous tissue preexposed to ultrahigh-frequency waves. A quantitative criterion is proposed for assessing the efficacy of the studied methods of exposure. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N o 6, pp. 669–672, June, 1995  相似文献   

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