制备高效价、高特异性的新型α-半乳糖苷酶的兔多抗,并鉴定该抗体的特异性。方法:用脆弱类杆菌来源的基因重组α-半乳糖苷酶(纯度大于90%)免疫新西兰大白兔,获得α-半乳糖苷酶的兔抗血清,并经HiTrap rProtein A柱纯化获得高纯度的抗体;用间接ELISA法检测抗体效价,Western印迹评价抗体的特异性。结果:通过免疫法得到了α-半乳糖苷酶的兔多克隆抗体血清,抗体效价达1∶1×106,经rProtein A柱纯化后获得了高效价、高纯度的抗体,Western印迹显示该抗体特异性地与新型α-半乳糖苷酶结合。结论:获得了新型α-半乳糖苷酶的高效价、高特异性的兔多克隆抗体,可用于血型转变过程中残留α-半乳糖苷酶含量的特异性检测。

Objective: To prepare high titer and high specific rabbit polyclonal antibody against a novel α-galac-tosidase from Bacteroides fragilis. Methods: The New Zealand rabbits were immunized with purified recombinant bacteria α-galactosidase(the purity>90%). Rabbit sera were purified by HiTrap rProtein A column, and its titer and specificity were detected by ELISA and Western blotting respectively Results: The purity of antibody protein was about 95%. And the titer of rabbit sera were 1∶1 × 106. Western blotting showed that the antibody reacted with α-galactosidase only. Conclusion: We had obtained high titer and high purity rabbit polyclonal antibody ofα-galactosidase. The antibody can be used in detecting the minimal amount of residual α-galactosidase involved in conversion of blood type B to O.

目的：分析和比较临床上采用酶联免疫吸附法(ELISA)以及化学发光法检测脐血中HIV-1/HIV-2抗体、梅毒抗体以及丙肝抗体的结果。方法：2011年8月-2014年8月收集脐血1147份，对其临床数据进行回顾性分析，分别采用酶联免疫吸附以及化学发光法对其中的HIV-1/HIV-2抗体、梅毒抗体和丙肝抗体进行检测，并对结果进行分析。结果：1147份脐血中酶联免疫吸附方法检测 HIV-1/HIV-2抗体、梅毒抗体和丙肝抗体阳性率分别为0.61%(7/1147)、0.87%(10/1147)、1.13%(13/1147)，而化学发光法检测HIV-1/HIV-2抗体、梅毒抗体和丙肝抗体阳性率分别为0.78%(9/1147)、0.96%(11/1147)、1.39%(16/1147)，并且将购买的抗体标准品进行梯度稀释。检测结果表明，3种抗体稀释到10 pg/mL，化学发光法检测3组抗体均为阳性，酶联免疫吸附方法仅梅毒抗体为阳性，而HIV-1/HIV-2抗体、丙肝抗体的检测结果均为阴性。结论：本研究结果表明，临床上采用化学发光发对脐血中HIV-1/HIV-2抗体、梅毒抗体和丙肝抗体检测的灵敏性高于ELISA方法，因此在临床检测过程中建议采用化学发光发进行检测。

Objective:To analyze and compare the difference of enzyme linked immunosorbent assay(ELISA) and chemiluminescence method in detection of HIV-1/HIV-2 antibody,syphilis antibody and hepatitis C antibody on umbilical cord blood.Methods:1 147 units of cord blood were collected in our hospital from August 2011 to August 2014.The clinical data were retrospectively analyzed,using enzyme linked immunosorbent assay and chemiluminescence method respectively to detecte HIV-1/HIV-2 antibody,the syphilis antibody and hepatitis C antibody,then we analyzed the results.Results:Among those 1147 units of cord blood,the positive rate of HIV-1/HIV-2 antibody,syphilis antibody and HCV antibody were 0.61%(7/1147),0.87%(10/1147) and 1.13% (13/1147) respectively by using enzyme linked immunosorbent assay detection method,while by chemiluminescence detection the positive rates of HIV-1/HIV-2 antibody and syphilis antibody and hepatitis C antibody were 0.78%(9/1147),0.96%(11/1147) and 1.39%(16/1147) r

目的：探讨就诊人群中抗环瓜氨酸肽（CCP）抗体、抗角蛋白抗体（AKA）、抗 RA33抗体、磷酸6葡萄糖异构酶（GPI）、类风湿因子（RF）等类风湿关节炎相关抗体的阳性分布趋势，及其在不同疾病中的分布与临床应用价值。方法用固相酶联免疫吸附法（ELISA）检测抗 CCP 抗体、抗 RA33抗体、磷酸6葡萄糖异构酶（GPI）、用间接免疫荧光法检测 AKA，用免疫比浊法检测 RF。结合患者临床资料分析这些自身抗体在不同年龄、性别和疾病中的阳性分布趋势及临床应用价值。结果抗 CCP 抗体、AKA、抗 RA33抗体、GPI、RF 例就诊人群的阳性率分别为26．22％、6．65％、7．13％、41．19％、32．95％；女性中抗 RA33抗体阳性率高于男性，男性抗 CCP 抗体阳性率高于女性（P ＜0．05）。在 RA 患者组各种抗体阳性率明显高于其他组，且抗体滴度也明显高于其他组。结论抗 CCP 抗体、AKA、抗 RA33抗体、GPI、RF 主要见于类风湿关节炎患者，但是也可出现在其他疾病中，但滴度较低。因此在临床诊疗过程需综合分析，应避免片面根据自身抗体检查结果所致误诊。

Objective To explore the positive distribution of anti-cyclic citrullinated peptide(CCP)antibody, anti-keratin antibody(AKA),anti-RA33 antibody,glucose-6-phosphate isomerase(GPI),rheumatoid factors(RF)in population with rheumatoid arthritis,and to evaluate the association between these distributions and disease diagno-sis.Methods Anti-CCP antibody,anti-RA33 antibody,and GPI were measured by solid enzyme-linked immunosor-bent assay(ELISA).AKA was detected by indirect immunofluorescence assay.RF was determined by immunoturbidi-metric assay.The positive distribution of these autoantibodies in population of different ages,genders and diseases, and their clinical significance were investigated in combination with the clinical data of the patients.Results The pos-itive rate of Anti-CCP antibody,AKA,anti-RA33 antibody,GPI,and RF:26.22%,6.65%,7.13%,41.19%, 32.95%,respectively.The positive rate of anti-RA33 antibodywas higher in female than in male.The positive rate of patients with r

目的 目前有多种方法可以用于结核性胸腔积液的诊断.本研究比较了IFN-γ释放试验(IGRAs)、腺苷脱氨酶活性检测(ADA)和结核抗体检测(TB-ab)三种方法.方法对63例胸腔积液患者分别使用IFN-γ释放试验、腺苷脱氨酶活性检测和结核抗体检测三种方法进行鉴别诊断.IFN-γ释放试验、腺苷脱氨酶活性检测和结核抗体检测均根据厂家提供的方案进行.结果 三种方法都有一定的诊断意义,但在效果上有差别.其中,结核抗体检测敏感性和特异性最低.尽管腺苷脱氨酶活性检测的特异性较高,但其仅仅检测到58.14％的患者,敏感性较低.IFN-γ释放试验检测到了79.07％的患者,并且具有较高的特异性.结论IFN-γ释放试验检测具有比腺苷脱氨酶活性检测和结核抗体检测更好的效果,推荐在结核性胸腔积液的诊断中使用IFN-γ释放试验.

Objective To study the effect of interferon-gamma release assays (IGRAs) adenosine deaminase activity test (ADA) and tuberculosis antibody test (TB-ab) in diagnosis of tuberculous pleurisy.Materials Sixty-three patients with pleurisy were tested for IGRAs,ADA and tuberculosis antibody.Samples of blood were tested.IGRAs,ADA and tuberculosis antibody were diagnosed according to manufacturer'' s instructions.Results Either of the method could detect several patients with tuberculous pleurisy.Tuberculosis antibody showed lowest sensitivity and specificity.In our experiments,ADA test only detected 58.14％ of patients,although specificity can be high.We detected highest percentage of tuberculous pleurisy (79.07％) in blood IGRAs,with a specificity of 70％.Conclusion Blood IGRAs are more effective than blood ADA and tuberculosis antibody test,and it is recommend that IGRAs should be used in diagnosis of tuberculous pleurisy.

目的：探讨心肌酶、肌钙蛋白Ⅰ（ cTnⅠ）、肺炎支原体抗体检测在EV71型重症手足口病中的临床意义。方法选取2012年5月～2014年5月收治的215例EV71型重症手足口病患儿为观察组，同时选取普通EV71型手足口病病例120例作为对照组，分别检测两组血清心肌酶、cTnⅠ的水平，肺炎支原体IgM抗体，并分析观察组患儿治疗前后心肌酶、cTnⅠ的变化情况。结果观察组心肌酶、cTnⅠ高于对照组，差异具有统计学意义；肺炎支原体IgM抗体阳性率高于对照组；观察组、对照组患儿治疗后心肌酶、cTnⅠ水平较治疗前明显下降，差异具有统计学意义。结论手足口病病程中心肌较易受到侵犯，重症病例心肌酶、cTnⅠ升高更为明显；观察组和对照组相比较，肺炎支原体抗体的阳性率显著高于对照组，肺炎支原体感染可能为导致手足口病加重的一个诱因，手足口病心肌酶、cTnⅠ、肺炎支原体抗体检测的临床意义重大，可作为重症手足口病患儿临床检测的常规指标。

[ ABSTRACT] Objective To investigate the clinical significance of testing myocardial enzyme,troponinⅠand antibody of mycoplas-monia( MP-IgM) in children with severe hand-foot-mouth disease.Methods 215 cases of EV71 type with severe hand-foot-mouth disease from may 2012 to may 2014 in the Weifang people''s hospital were selected.As the observation group and 120 cases of regular hand-foot-mouth disease were chosen as the control group;the level of serum myocardial enzyme,cTnⅠ,anti-mycoplasmonia in the two groups were tested,the indications in the observation group before and after the treatment were analyzed.Results The level of myocardial enzyme,troponinⅠand the positive rate of anti-MP-IgM in the observation group were higher than those in the control group and the differences were statistical significant ( P<0.01);the level of myocardial enzyme,troponin Ⅰin observation group after the treatment were lower than those before the treatment, the differences were statistical