对流免疫电泳法鉴定蚊胃血血源的研究

用对流免疫电泳技术(电泳法),以全血清为抗原,制备3种已知抗体血清检测未知抗原鉴定蚊虫胃血。并用环状沉淀试验法(沉淀法)比较观察不同抗血清及抗原浓度、反应时间与温度的实验。结果证明沉淀法的敏感性比电泳法高约2～3倍。两法同时检测中华按蚊及菲律宾按蚊胃血标本,结果两法牛血阳性率均为100％,但沉淀法尚出现30.43％及17.24％牛、猪血阳性反应,而电泳法则无;当沉淀法抗血清稀释为1:32时,两法反应结果一致。电泳法操作简便,可用于蚊胃血标本的鉴定。     

定量检测内毒素的火箭电泳鲎试验法的建立及评价

用亚洲鲎血细胞溶解物中凝固蛋白原的抗血清建立的火箭电泳鲎试验法,适用于定量检测血中内毒素。该实验观察了标示灵敏度不同的鲎试剂与内毒素反应的曲线,以及保温时间和反应体积比对检测灵敏度的影响。采用氨甲酰化抗体免疫电泳系统和高压电泳技术,可缩短电泳时间2h,标示灵敏度为1.0Eu/ml、0.5Eu/ml和0.25Eu/ml的鲎试剂在本法的检测低限分别为0.06Eu/ml、0.03Eu/ml和0.007Eu/ml。检测不受样品色素或混浊的干扰。     

应用PEG沉淀试验测定囊虫病人血清中抗体的初步研究

用适宜浓度的PEG将囊虫病人血清浓缩,达到提高血清中特异性抗体浓度的作用,从而提高检测的敏感性。该法用于囊虫病的诊断,阳性附合率83.33%.特异性较理想。     

实验教学中颗粒性抗原与可溶性抗原制备抗血清的方法及其应用

目的比较颗粒性抗原与可溶性抗原制备抗血清的效价,寻求获取符合实验教学需要的高效价抗血清的较佳方法。方法牛血清白蛋白（Bovine Serum Albumin,BSA）作为可溶性抗原,伤寒“H”菌液作为颗粒性抗原,采用1周间隔的免疫程序,以BSA和伤寒杆菌“H”菌液免疫家兔,用对流免疫电泳和双向琼脂扩散试验、肥达试验检测并比较其血清抗体效价。结果浓度为7×109个/ml伤寒杆菌“H”菌液免疫家兔后获取的伤寒“H”抗血清几何平均滴度（geometry mean titer,GMT）为1∶9051.07,浓度为5mg/ml BSA免疫家兔后测得抗血清GMT分别为1∶3.668和1∶6.17。结论颗粒性抗原与可溶性抗原1周间隔免疫程序制备的抗血清可获取符合实验教学的抗血清效价。     

Serological cross-reactivity of porcine reference antisera to Mycoplasma hyopneumoniae, M. flocculare, M. hyorhinis and M. hyosynoviae indicated by the enzyme-linked immunosorbent assay, complement fixation and indirect hemagglutination tests

The enzyme-linked immunosorbent assay (ELISA) indicated significant cross-reactivity between the antigens of Mycoplasma hyopneumoniae ( HyoP ) and M. flocculare (Floc), another porcine mycoplasma of wide distribution but uncertain pathogenic significance, when porcine antisera of each specificity were tested against HyoP antigen. The titers of the anti-Floc sera ranged from threefold to 13-fold less than the titer of the anti- HyoP reference serum at different times after immunization. These values ranged from onefold less than to fourfold greater than the minimal positive titer of 80. The antisera to the other porcine mycoplasmal antigens [i.e. M. hyorhinis ( HyoR ) and M. hyosynoviae ( HyoS )] reacted less strongly to HyoP antigen but titers only slightly less than to slightly greater than the minimal positive titer were noted for some sera. Cross-reactivity was also detected by the complement fixation test, although the titers for this test were generally lower than for the ELISA, presumably reflecting lower sensitivity of the complement fixation test. Positive indirect hemagglutination titers to HyoP antigen were also observed for both anti-Floc sera obtained at one or more times during the immune response. With two exceptions (one anti- HyoR serum with a complement fixation titer of 16 and one anti- HyoR serum with an indirect hemagglutination titer of 10), none of the anti- HyoR or anti- HyoS sera had detectable indirect hemagglutination or complement fixation titers to HyoP antigen at any time after immunization. The levels of cross-reactivity detected by the complement fixation test and indirect hemagglutination and, especially, the ELISA would be of significance for the development of any practical sero-diagnostic test for mycoplasmal pneumonia of swine.     

不同免疫程序制备兔抗牛血清白蛋白抗血清的效价比较

目的比较两种免疫程序制备牛血清白蛋白（BSA）抗血清的效价,以获取能满足实验教学需要的高效价可溶性抗血清。方法采用2天短期间隔免疫程序和1周间隔免疫程序,以牛血清白蛋白（BSA）免疫家兔,对流免疫电泳和双向琼脂扩散试验检测并比较其血清抗体效价。结果BSA浓度在5mg／mL时,双向琼脂扩散试验和对流免疫电泳测得的2天间隔免疫程序所制备的BSA抗血清几何平均滴度（GMT）分别为1：6．50和1：4．2,1周间隔免疫程序的BSA抗血清GMT分别为1：10．56和1：8．00。结论1周间隔免疫程序所制备的牛BSA抗血清效价高于2天短周期间隔的免疫程序所制备抗血清效价,BSA短周期免疫家兔可获取符合教学实验要求的抗血清效价。     

Sensitivity and specificity of an enzyme-linked immunosorbent assay for the detection of infectious bovine rhinotracheitis viral antibody in cattle

An enzyme-linked immunosorbent assay was developed to detect bovine serum antibody to infectious bovine rhinotracheitis virus. The specificity of this assay in 304 bovine sera, collected from an infectious bovine rhinotracheitis virus-free herd, was 100%; in sera from 62 cattle inoculated with an intranasal vaccine, its diagnostic sensitivity was 27.4% at one month and 100% at six months, postvaccination. In 303 bovine sera with standard serum neutralizing antibody titers of greater than or equal to 1:2 it showed 100% sensitivity; and in 463 random diagnostic samples, comparative tests indicated that enzyme-linked immunosorbent assay detected more seropositive animals (61.6%) than the standard serum neutralizing test (49.9%). The enzyme-linked immunosorbent assay method was considered to be technically superior as a routine diagnostic test for the detection of infectious bovine rhinotracheitis viral antibody in bovine sera.     

卵骒癌免诊的初步研究

Ovarian tumor associated antigen was isolated from human serous cystadenocarcinomas of ovary and purified by using affinity chromatography columns coupled to anti-normal female serum and anti-normal ovarian tissue gamma globulin. Antiserum was prepared from New Zealand white rabbits. The antiserum was absorbed with normal female serum and extracts of normal ovarian tissue thoroughly. The absorbed antiserum was appraised by immunodiffusion, immunoelectrophoresis and counter-immunoelectrophoresis with intensifiers. The absorbed antiserum has a specific immune precipitin with extracts of ovarian tumor tissue. Counter-immunoelectrophoresis with intensifiers was used to detect the ovarian tumor associated antigen in the tissue and serum of various patients. Twenty-eight of 40 ovarian tumor extracts and 1 of 5 cervical carcinoma extracts showed the presence of the cancer antigen. 14/40 were positive in the sera of ovarian tumor patients and 6/10 positive in the sera of ovarian serous cystadenocarcinoma patients; 1/5 positive in the sera of cervical carcinoma patients and 2/10 positive in the sera of ovarian cysts patients. No positive result for the sera of normal females was noted. The specificity, sensitivity and accuracy of the method were 94.7%, 42.3% and 83.3%, respectively.     

肌钙蛋白I（cTnI）定量检测诊断AMI的评价

目的评价肌钙蛋白I（cTnI）定性检测对诊断急性心肌梗死（AMI）的应用价值。方法对162例入院诊断急性心梗患者,分别作肌钙蛋白I定量检测以及冠状动脉造影,计算cTnI检测诊断心肌梗死的灵敏度和特异性,组间差异采用χ2检验。结果cTnI定量检测的灵敏度为91.3%,特异性为100%,诊断AMI2组有显著差异性。结论cTnI定量检测临床诊断AMI不能取代冠脉造影。cTnI诊断AMI具有高灵敏度,高特异性的特点,是临床诊断AMI有效指标。     

Reactivity of human preformed natural antibodies with various porcine pancreatic cells

Pancreatic islet transplantation is a promisingtherapeutic approach for the treatment of insulin- de-pendent diabetes mellitus( IDDM) [1] .Because of se-vere shortage of human organs,the pig appears to bethe most likely donor species for xenotransplantationinto humans for a variety of reasons[2 ] .However,successful organ xenotransplantation requireselimina-tion of preformed natural antibodies( PNAbs) re-sponsible forhyperacute graftrejection.Itis notwellknown whether human PNAbs to the po…     

制备抗人血清白蛋白抗体的简单高效方法

以简单的酸乙醇法纯化人血清白蛋白(HSA)作为抗原,采用一个高效、经济、快速的免疫方案免疫家兔,得到高效价的抗血清。最后利用ProteinA-Sepharose 4B亲和层析法纯化IgG抗体。     

含丹参酮ⅡA 磺酸钠小鼠血清的制备及其对PCASM Cs的BKCa作用的初探

目的：探讨一种将中药血清药理学和电生理学技术结合的方法，并将该方法运用到中药丹参的有效成分丹参酮ⅡA磺酸钠（DS‐201）的舒血管机制研究。方法采用醇提法提取丹参有效成分，将该提取液给予小鼠灌胃，经眼球采血得到小鼠血清，用高效液相色谱法（HPLC）测定小鼠血清中DS‐201的含量，并将该含药血清应用于单通道膜片钳实验，研究其对猪冠状动脉平滑肌细胞（PCASMCs）大电导钙激活钾通道（BKCa ）的作用。结果含DS‐201血清浓度线性范围是0．73～1．91μg／mL （r＝0．9977），回收率是99．85％～101．47％，血药浓度为（7．32±4．25）μg／mL ；单通道膜片钳实验结果显示小鼠含DS‐201血清对PCASMCs的BKCa有激活作用，但差异无统计学意义（P＞0．05）。结论该研究建立的提取丹参有效成分丹参酮的醇提法简单、实用、可靠，且测定DS‐201含量的HPLC方法精确；应选取高质量丹参和（或）提高丹参在体内的生物利用度，才能更好地将血清药理学和电生理学技术结合在一起。     

血清EBV抗体检测联合99mTC-MIBI显像和MRI在诊断鼻咽癌放射治疗后复发中的价值

目的:探讨血清EBV抗体检测联合^99mTc-MIBI显像和MRI在诊断鼻咽癌放射治疗后复发中的价值。方法:60例鼻咽癌放射治疗后患者依据病理组织活检和临床随访复发21例,随访中均行血清EBV抗体检测、^99mTc-MIBI显像显像和MRI检查。分别评价三者对鼻咽癌放疗后复发检出的灵敏度、特异性和准确度进行比较。结果:联合3种检查方法诊断鼻咽癌放疗后复发的灵敏度、特异性和准确率分别为100%、97.44%、和98.33%,其中灵敏度与单一检查方法比较差异无统计学意义;特异性与亲肿瘤显像比较差异无统计学意义,但与血清EBV抗体检测和MRI检查差异均具有统计学意义;准确率与单一检查方法差异均有统计学意义。结论:联合血清EBV抗体检测、^99mTc-MIBI显像和MRI在诊断鼻咽癌放射治疗后复发中具有较高的价值,三种诊断方法有各自的优缺点,三者结合起来应用,可明显提高诊断鼻咽癌放射治疗后复发的准确性。     

用猪血清Clq提取物检测循环性免疫复合物

我们采用Yonemasu优球沉淀及Bing亲和层析法,并附加SPA菌体吸收IgG的步骤,从猪血清分离Clq。经热灭活,化学灭活,4％SDS-PAGE分析证明不仅在Clq提取物中而且在高度提纯的猪IgM中均含有IgM-RF。用猪Clq作固相ELIZA,试验的敏感范围是10-1000ngHAG/mL。38例正常人血清的测定值是2.06±1.52μgHAG/Ml(m±SD)。测13例SLE及16例RA病人血清,其值分别为31％及62.5％,超过m+2SD。用猪Clq测CIG可取得满意结果。     

分光光度法对肿瘤特异生长因子的灵敏度及检测极限

目的：用721可见分光光度法对随机混合血清的肿瘤特异生长因子(TSGF)值进行测定。考察该方法的灵敏度及检测极限。方法：严格按福建新大陆生物技术有限公司开发的TSGF联合检测试剂盒使用说明书进行。该试剂在特定条件下，与血清中的多种肿瘤相关物质产生显色反应。结果：721可见分光光度法的灵敏度为k=0．00586，检测极限为1．52U／ml。结论：721分光光度法对ISGF有较高的灵敏度和较理想的检测极限。该方法所需设备简单，操作简便快速，是基层医院及预防中心对肿瘤进行普查及筛选的理想方法，可推广应用。     

牛血清白蛋白的抗血清在免疫学沉淀反应实验中的应用

目的 :制备牛血清白蛋白 (BSA)抗血清 ,以探讨获取高效价抗血清的方法。方法 :用BSA配制一定的浓度 ,免疫家兔 ,以制备兔抗 -BSA血清。利用免疫电泳、双向琼脂扩散试验分析和测定抗血清的效价。结果 :双向琼脂扩散在BSA 0 . 5g/mL的浓度时 ,测得抗血清效价为 1∶32 ,对流免疫电泳结果显示BSA 0. 5g/mL与抗BSA在 1∶16时形成一条清晰可见的沉淀线。结论 :BSA免疫家兔可获取效价较高的抗血清。     

Development of an immunoelectroosmophoresis test for the detection and typing of antibodies to vesicular stomatitis viruses

This study was conducted to develop a rapid test to detect type specific antibodies to various strains of vesicular stomatitis virus. Glycoprotein was extracted from purified vesicular stomatitis virus-Indiana 3 and used as antigen in an immunoelectroosmophoresis test (counter immunoelectrophoresis) to test reactivity with homologous hyperimmune serum and antisera to vesicular stomatitis virus-New Jersey, Indiana 1 and Indiana 2. A reaction was only detected with the homologous antiserum. A simpler preparation of antigen, a detergent extract of infected cells was also evaluated and shown to have less specificity in reactions with hyperimmune sera. This infected cell extract was, however, useful in detecting homologous antibodies in convalescent sera (natural vesicular stomatitis virus-New Jersey infection) and no reactions were detected with infected cell extracts of vesicular stomatitis virus-Indiana 1, 2 and 3.     

纳米金增敏 cTnI 金标免疫层析试验研究

目的：研究一种纳米金增强心肌肌钙蛋白I（cTnI）金标免疫层析试验灵敏度的试验方法。方法以血清cTnI金标免疫层析试验为基础,以改性溶液做预处理,盐酸羟胺还原氯金酸制备纳米金颗粒,使金标抗原抗体反应复合物的颗粒增大,金标处颜色加深,显色信号增强,提高检测灵敏度,并以此做批内和日间重复性检测。结果盐酸羟胺还原氯金酸增敏cTnI金标免疫层析试验,较增敏前检测灵敏度提高16倍,在10～15 min内可完成,批内和日间重复性较好,试验结果快速、稳定、可靠。结论盐酸羟胺还原氯金酸制备纳米金颗粒在一定改性溶液环境下,对人血清cTnI金标免疫层析试验有一定增敏效果,重复性较好,结果稳定可靠,可为适时、快速、灵敏的现场检测方法提供一定技术支持。     

子宫内膜异位性疾病非手术法诊断的可行性探讨

目的评估非手术方法诊断子宫内膜异位性疾病的临床可行性。方法回顾性分析因痛经、不孕、盆腔包块结合超声,血清CA125检查初步诊断为子宫内膜异位性疾病而住院行手术治疗的患者179例。计算痛经、妇科检查阳性体征、超声和血清CA125指标单独及两项联合诊断内异性疾病的敏感性、特异性、阳性预测值、阴性预测值。结果各单指标诊断内异症性疾病的灵敏性、特异性均较低,两指标联合的平行试验可将敏感性提高至85.54%,两指标联合系列实验特异性可提高至85.90%。痛经、妇科检查、超声、血清CA125,诊断子宫型内异性疾病的阳性率分别为79.07%、7.44%、72.09%、76.74%;诊断卵巢型内异症的阳性率分别为73.68%、60.53%、76.32%、71.05%;子宫型合并卵巢型内异症的阳性率除妇科检查外均高于单一型内异症。结论痛经结合临床诊治、血清学指标的非手术法诊断内异症性疾病是可行的,各指标联合诊断可提高内异性疾病的临床诊断价值,除超声指标外其他指标诊断子宫型内异症的价值较卵巢型内异症高。     

人促黄体生成素光激化学发光免疫分析法的建立

目的采用光激化学发光免疫分析技术建立快速定量检测人促黄体生成素(hLH)的方法。方法用2株配对的hLH单克隆抗体,1株hLH单克隆抗体包被受体微球,另1株hLH单克隆抗体先用生物素标记,再与链霉亲合素的供体微球共同组成人促黄体生成素光激化学发光免疫分析试剂,进而优化反应体系并对试剂的各项性能指标进行评价。结果自制hLH试剂分析灵敏度为0.164 U/L,线性测量范围为0.164～135 U/L,分析内和分析间的精密度分别为4.3%～6.1%和7.4%～8.1%,均低于10%,与hTSH、hFSH和hCG无明显交叉反应,158份临床血清样本用本试剂与罗氏电化学发光检测试剂盒平行检测,对所测数值采用配对t检验分析,结果显示两种方法检测结果差异无统计学意义(t=-1.468,P=0.07)。结论自制hLH光激化学发光免疫分析试剂各项指标均能达到临床要求,有望替代国外同类产品。