血清结核杆菌抗体检测在呼吸道感染鉴别诊断中的意义

为探讨血清特异性结核抗体的辅助诊断价值和加强对小儿肺炎的鉴别诊断，采用酶联免疫吸附试验（ＥＬＩＳＡ）法，对１５７７例肺炎患儿、６８５例健康儿童及１１１例支气管淋巴结结核患儿进行了血清特异性结核抗体检测，用纯蛋白衍生物皮试作对照，肺炎组还作了结核杆菌ＰＣＲ检查。结果表明，结核组患儿的结核抗体阳性８４例（７５．７％），健康组儿童的结核抗体阳性２４例（３．５％），肺炎组的结核抗体阳性２６２例（１６．６％），肺炎组与健康组比较，ｕ＝８．６３，Ｐ＜０．０１。符合儿童结核诊断者３９例，占受检病例的２．５％，占结核抗体阳性病例的１４．９％。ＥＬＩＳＡ法的敏感性为０．７５７、特异性为０．９６５、符合率为０．９３６、阳性预测值为０．７７８、阴性预测值为０．９６１、精确性为０．７２２，９５％可信限为０．６１２～０．８３２之间。提示检测结核抗体有重要辅助诊断价值；肺炎患儿的结核感染率远高于健康组，故应高度警惕，在肺炎患儿中发现结核病患儿，并将其作为重点防治对象。     

Hepatitis C virus infection and related liver disease in children of mothers with antibodies to the virus

Objective: To evaluate the clinical, biochemical, and virologic features associated with hepatitis C virus (HCV) infection acquired early in life from mothers with antibodies to HCV (anti-HCV).Study design: Multicenter prospective-retrospective study in Italian children.Patients: Two groups of children were investigated. Group 1 included 14 infants, born to mothers with anti-HCV but without human immunodeficiency virus infection, who became seropositive for HCV RNA during the first year of life and were thus considered infected. Group 2 included 16 children with chronic hepatitis C, aged 1 ½ to 14 years, whose mothers were the unique potential source of infection. Both groups were followed for 12 to 48 months.Methods: Alanine transaminase (ALT), anti-HCV, and HCV RNA were investigated by the polymerase chain reaction on entry to the study and during follow-up.Results: All children in group 1 had anti-HCV throughout follow-up, and all had ALT abnormalities, ranging from 1.5 to 10.5 times the normal value during the first 12 months. During further follow-up, 5 of 10 children had HCV RNA with abnormal ALT values, 3 had a return to normal of the ALT values but continued to have viremia, and 2 eventually had normal ALT values and clearance of HCV RNA. Of the 16 children in group 2, all were free of symptoms and 62% had only slight ALT elevations; 7 who underwent liver biopsy had histologic features of minimal or moderate hepatitis.Conclusions: HCV infection acquired early in life from mothers with anti-HCV is usually associated with biochemical features of liver damage during the first 12 months of life. Progression to chronicity seems to occur in the majority of cases, although HCV-associated liver disease is likely to be mild throughout infancy and childhood. (J Pediatr 1997;130:990-3)     

抗人喉癌单克隆抗体对荷瘤裸鼠放射免疫定位显像的观察

目的用喉癌裸鼠模型研究抗人喉癌单克隆抗体的免疫导向作用，为诊断和治疗提供依据和方向。方法用高效碘标法标记三株混合抗人喉癌单克隆抗体（ＭｃＡｂＬＣ）经腹腔注入人喉癌裸鼠模型体内，用单光子发射计算机断层（ｓｉｎｇｌｅｐｈｏｔｏｎｅｍｉｓｓｉｏｎｃｏｍｐｕｔｅｄｔｏｍｏｇｒａｐｈｙ，ＥＣＴ）扫描显像并测定肿瘤、血液和组织单位质量浓集的放射性占注入总放射性的百分比（％ＩＤ／ｇ）和肿瘤与主要器官组织的放射性比值（Ｔ／ＮＴ）。结果单克隆抗体在一定时限内能够选择性的浓集在移植瘤组织内，９６ｈ可获得满意的肿瘤显像，实验组肿瘤组织的％ＩＤ／ｇ为１８．８±０．６１，对照组肿瘤组织的％ＩＤ／ｇ为４．４６±０．８２，两组比较差异有显著性（Ｐ＜０．０１，ｔ＝３４．９７）。结论用放射性核素标记抗人喉癌单克隆抗体在活体内能选择性地与人喉癌细胞结合，用ＥＣＴ扫描可以识别肿瘤所在的部位和大小，对肿瘤病灶及转移病灶的定位诊断有一定的实用价值。     

MG_7、MGd_1单克隆抗体在卵巢粘液上皮肿瘤中的表达及对患者预后的意义

目的：探讨抗胃癌单克隆抗体ＭＧ７、ＭＧｄ１在卵巢粘液上皮肿瘤（ＯＭＥＴ）组织中的表达与临床资料和患者预后的关系。方法：应用免疫组化ＳＰ法检测９３份ＯＭＥＴ组织中ＭＧ７、ＭＧｄ１的表达，并对１４ａ经手术切除、病理学检查证实的ＯＭＥＴ患者的临床资料进行患者预后分析。结果：ＭＧ７、ＭＧｄ１定位于ＯＭＥＴ细胞膜及附近的胞浆，ＭＧ７在良性、交界性、恶性组表达率分别为０、８０．６５％、１００％，ＭＧｄ１分别为２６．４７％、９６．７７％、１００％。３组间差异有显著性（Ｐ＜０．０５）。恶性组ＭＧ７过度表达率比ＭＧｄ１者高１４．２８％。将本组ＯＭＥＴ的临床资料、ＭＧ７和ＭＧｄ１染色结果分别经ＣＯＸ风险模型分析，发现期别、肿瘤直径和ＭＧ７染色指标是影响患者预后的主要因素。ＯＭＥＴＭＧ７表达（—）、（＋）、（）和（）组的患者５ａ生存率分别为１００％、７２．３７％、２７．９％和１３．１３％，ＭＧｄ１分别为（—）１００％、（＋）７５．０５％、（）３１．４７％、（）０．００％。结论：卵巢癌与胃癌组织间存在共同抗原性，具有同源性。ＭＧ７、ＭＧｄ１对ＯＭＥＴ的诊断有一定的特异性和敏感性，与临床病理分级、分类及预后有关，为ＯＭＥＴ的?     

The sternocleidomastoid test: an in vivo assay to investigate botulinum toxin antibody formation in humans

In a small number of patients treated with botulinum toxin (BT) antibody (Ab) formation occurs. BT Ab can be detected by the mouse protection assay (MPA) or by the mouse diaphragm assay (MDA). Both methods, however, have major drawbacks. We tested a method for detecting BT Ab which measures the BT-induced reduction in the electromyographic amplitude of the mean maximal voluntary activation (M-EMG) of the sternocleidomastoid muscle. The M-EMG reduction was compared in 17 patients with cervical dystonia and secondary BT therapy failure to the M-EMG reduction previously measured in controls. Values more than 2 SD below the mean of controls were considered abnormal. Six patients showed BT Ab on the MPA and MDA; all of these had abnormal M-EMG reductions. Eleven patients showed no BT Ab on MPA and MDA testing; in ten of these the M-EMG reduction was normal, and in one it was pathological, but MDA testing later changed to positive under continued BT therapy. The sternocleidomastoid test is easy to perform and produces quantitative results. Since its sensitivity and specificity are at least as good as those of the MDA and the MPA, it can replace them.     

肿瘤相关抗原抗体联合检测在肺癌早期诊断中的价值评价

目的：对肿瘤相关抗原（tumor—associated antigen，TAA）抗体联合检测用于肺癌诊断的价值进行评价。方法：利用酶联免疫吸附试验（ELISA）检测肺癌患者血清和正常血清中8种TAA抗体，并进一步利用流行病学方法对各抗原检测结果进行评价。结果：每种抗原检测结果单独进行判断时，灵敏度都偏低。对8种TAA进行不同的组合，结果随着抗原种类的增多，灵敏度随之增加，8种抗原联合检测的灵敏度达到了64．3％，特异度达到了86．2％，阳性预测值88．7％，阴性预测值58．8％，提示8种抗原组合检测肺癌时大大提高了诊断的质量；阳性似然比4．66，说明8种抗原联合检测肺癌的临床诊断价值较高；kappa值0．46，提示该试验诊断结果与真实值之间中度一致。结论：利用8种TAA抗体联合检测肺癌，具有较高的真实性，为现场高危人群的筛检和临床中肺癌的早期诊断提供了一种新方法。     

抗人间充质干细胞单克隆抗体的制备及生物学特性研究

目的:制备抗人间充质干细胞(hM SC s)的单克隆抗体,为鉴定、分选hM SC s奠定基础。方法:以多供体hM SC s免疫BALB/C小鼠,采用杂交瘤技术制备单克隆抗体,并以间接免疫荧光法、免疫组织化学法和流式细胞术等检测单克隆抗体在细胞及组织中的表达。结果:建立了5个能分泌抗人间充质干细胞的单克隆抗体杂交瘤细胞株,该组单抗能与人间充质干细胞特异性结合,而对所检测的人间充质和非间充质来源的组织和细胞均未见交叉反应,5个单抗的相对亲和力分别为:ZUB1 1×106、ZUB4 1×105、ZUC 3 1×106、ZUE 12 1×105、ZUF10 1×105;表位分析:ZUB1、ZUE 12、ZUF10针对相同的表位,ZUC 3和ZUB4则分别针对不同的表位。流式细胞分析显示分离培养的hM SC s阳性表达率为ZUB1 87.39%、ZUB4 88.07%、ZUC 3 88.12%、ZUE 12 69.89%和ZUF10 83.67%。结论:5株杂交瘤分泌的单克隆抗体均能与人间充质干细胞发生特异性结合,为深入研究hM SC s的生物学功能奠定了基础。     

酶联免疫吸附试验检测血清抗中性粒细胞胞质抗体方法的建立与临床应用

目的建立总抗原酶联免疫吸附试验(ELISA)检测血清抗中性粒细胞胞质抗体(ANCA)的方法,初步探讨其在ANCA相关血管炎中的表达及临床应用价值。方法从人的中性粒细胞中提取总抗原,用棋盘滴定法选择ELISA测定的最佳条件,确定临界值。测定系统性红斑狼疮(SLE)、肾病、血管炎、类风湿关节炎(RA)等患者血清总ANCA,并与间接免疫荧光(IIF)法、特异性髓过氧化物酶(MPO)-ANCA及蛋白酶3(PR3)-ANCA进行分析比较。结果总抗原ELISA与IIF法检测肾病、SLE、血管炎及RA患者血清ANCA阳性率比较差异均无统计学意义(P>0.05)。抗原特异性ELISA、IIF法检测肾病、血管炎、RA患者血清ANCA阳性率比较差异均无统计学意义(P>0.05),而SLE差异有统计学意义(P<0.05)。结论ELISA检测ANCA可行且具有广泛的推广应用前景。     

抗脂多糖单链噬菌体抗体在大肠埃希菌中的表达及筛选

目的研究小鼠鼠源性的抗脂多糖(lipopolysaccharide, LPS)单链可变区片段(single-chain fragment variable, ScFv),即在大肠埃希菌TG1中的表达,并筛选出对LPS具有较高亲和力的ScFv. 方法将预先构建好的有效库容量为4.75×106的噬菌体抗体库接种至SOBAG培养基上,隔夜培养后扩容至约1×1010;严格控制lac启动子的表达,仅当加入M13KO7辅助噬菌体后才使ScFv以附着型的形式表达,然后利用LPS淘筛特异性的ScFv,富集的噬菌体阳性克隆重新感染TG1;最后随机挑选出190个菌落在96孔培养板分别培养单个含特异性ScFv的TG1菌落,经间接ELISA检测抗LPS的ScFv. 结果淘筛一轮过后即有3×104阳性菌落长出,190个菌落经间接ELISA检测有两个阳性克隆.结论成功地使鼠抗LPS ScFv在大肠埃希菌中表达,并从中筛选出了两株抗LPS ScFv.