抗CCP抗体、AKA及APF联合检测在类风湿关节炎诊断中的应用价值

目的观察抗环瓜氨酸肽(CCP)抗体、抗角蛋白抗体(AKA)及核周因子(APF)对类风湿性关节炎(RA)的诊断价值。方法选择RA患者110例(RA组)、其他自身免疫性疾病患者50例(非RA组)和健康体检者110例(对照组)为研究对象,采用间接免疫荧光法检测AKA、APF,用酶联免疫吸附试验(ELISA)检测抗CCP抗体。结果 RA组患者3项检测指标阳性率均明显高于非RA组和对照组,差异均有统计学意义(P<0.05);3项目串联试验的灵敏度和特异性分别为44.55%、99.38%,3项目并联试验的灵敏度和特异性分别为93.64%、85.63%。结论在RA的诊断中抗CCP抗体具有较高的特异性和灵敏度,3项目串联试验可以提高特异性,降低误诊率,3项目并联试验可以提高灵敏度,降低漏诊率,3项目联合检测指导临床对RA的诊断具有重要价值。

结核分枝杆菌TB—SA抗体检测在结核病诊断中价值的研究

目的研究结核分枝杆菌TB—SA抗体检测在结核病诊断中的价值。方法对1232例结核病、非结核病的其他呼吸系统疾患患者及健康志愿者同日寸进行抗酸杆菌涂片、培养、PPD皮试及结核分枝杆菌TB—SA抗体检测。结果①结核分枝杆菌TB—SA抗体检测菌阳肺结核的敏感性为75．1％；菌阴肺结核的敏感性为68．9％；肺外结核病的敏感性为71．2％；诊断结核病的总体敏感性为72．0％，特异性为82．1％。②TB—SA血清抗体检测OD405值并不与PPD值成线性关系，结核分枝杆菌TB—SA抗体检测对结核病人的诊断不受患者卡介苗反应的影响。结论结核分枝杆菌TB—SA抗体检测诊断结核病有较好的敏感性和特异性，是结核病诊断和鉴别诊断的可靠手段。

抗登革病毒单克隆抗体诊断盒的应用

应用４株Ｉ、Ⅱ、Ⅲ、Ⅳ型登革病毒特异性单克隆抗体组成的诊断试剂盒，用荧光抗体技术特异而敏感地鉴定了Ⅰ～Ⅳ型登革病毒原型株和２０个地方株的型别．并于１９９０－１９９３年间广州市及佛山市登革热流行时，对从病人血清分离的登革病毒准确而快速地进行型别鉴定。

IgA和IgG型抗中性粒细胞胞质抗体联合检测诊断韦格纳肉芽肿的价值

【目的】探讨IgA和IgG型抗中性粒细胞胞质抗体（ANCA）联合检测在韦格纳肉芽肿（WG）临床诊断中的应用价值。【方法】对28例临床诊断为wG及172例其他自身免疫性疾病患者血清，应用间接免疫荧光法检测ANCAIgA和IgG型抗体。【结果】wG病患者胞质型ANCA（cANCA）IgG型抗体和cANCAIgA型抗体阳性率最高，均显著高于其他自身免疫性疾病，且差异有显著性（P〈0．05）。联合检测ANCAIgG及IgA亚型抗体阳性率为89．3％（25／28），明显高于单独检测ANCAIgG或IgA亚型阳性率53．6％（15／28）和35．7％（10／28），且差异有显著性（P〈0．05）。【结论lIgA和IgG型抗中性粒细胞胞浆抗体联合检测有助于wG的早期准确诊断。

以合成肽为抗原建立ELISA试验检测鼻咽癌患者EB病毒抗体的研究

目的:以人工合成肽为抗原基质建立酶联免疫吸附试验,检测EB病毒抗体。方法:以人工合成肽作为抗原,建立ELISA试验,并检测93例鼻咽癌患者和106名正常人的血清。结果:以临界值0.259为阳性判断标准,93例鼻咽癌患者血清中,阳性63例,敏感度为67.7%;106名正常人血清中,阳性8名,特异度为92.5%。结论:所建立的以合成肽为抗原基质的EB病毒酶联免疫试验适用于鼻咽癌患者的筛查。

重组EBV-TK及DNase检测鼻咽癌血清相应抗体的比较研究

【目的】比较鼻咽癌(NPC)患者血清中EB病毒(EBV)胸苷激酶(TK)和DNA酶(DNase)抗体水平。【方法】用鼻咽癌血清中和Raji细胞产生的EBVDNase,抗酶率(AER)≥30%为EBVDNase抗体(EDAb)阳性。以大肠杆菌表达的重组EBVTK为抗原,用Westernblot方法检测鼻咽癌血清中的EBVTK抗体(ETAb),显现棕褐色条带者为ETAb阳性。【结果】55例鼻咽癌患者血清,EDAb阳性率94.5%,ETAb阳性率89.1%,EDAb+/ETAb+、EDAb-/ETAb+、EDAb+/ETAb-、EDAb-/ETAb-的检出率分别为87.3%、1.8%、7.3%和3.6%,同时检测此两项指标可使EBV相关抗原抗体的阳性率提高至96.4%。【结论】大多数鼻咽癌血清均同时具有EDAb和ETAb,检测这两个酶的抗体是鼻咽癌临床辅助诊断的二个重要血清学指标,联合应用意义更大。

抗环瓜氨酸肽抗体不同检测方法比较分析

目的比较分析电化学发光免疫(ECLIA)法与酶联免疫吸附(ELISA)法检测抗环瓜氨酸肽(CCP)抗体的相关性,并探讨其临床诊断效率和实用性。方法选择湖南省邵阳市中心医院2013年1月至2014年12月收治的类风湿关节炎患者210例作为观察组,另选择对照组178例,其中风湿性疾病患者71例,健康者107例。采用ECLIA法和ELISA法检测研究对象血清中抗CCP抗体,观察2种方法检测抗CCP抗体的诊断灵敏度和特异度。结果 ECLIA法检测抗CCP抗体的灵敏度为71.43%(150/210),特异度为98.31%(175/178);ELISA法检测抗CCP抗体的灵敏度为69.52%(146/210),特异度为96.63%(172/178)。2种方法测定388份血清标本,阳性147例,阴性230例。二者总体符合率为97.16%(377/388),差异有统计学意义(χ2=35.538 9,P<0.05)。结论ECLIA法检测抗CCP抗体对类风湿关节炎诊断的灵敏度、特异度与ELISA法具有较好的一致性,可在临床检验中推广应用。

p53自身抗体和Bmi-1自身抗体联合检测在肺癌诊断中的价值

目的评价p53自身抗体和Bmi-1自身抗体联合检测对肺癌的诊断效能。方法采用酶联免疫吸附试验检测92例肺癌患者和80例正常对照者血清中p53、Bmi-1自身抗体的表达水平，应用受试者工作特征曲线（ROC）分析两种自身抗体的联合诊断效能。结果肺癌患者血清p53、Bmi-1自身抗体的水平均高于正常对照，差异有统计学意义[（0．438±0．705）VS（0．045±0．035），Z=-7．667，P〈0．01；（0．117±0．061）VS（0．068±0．031）。Z=-7．179，P〈0．01]。血清p53、Bmi-1自身抗体在肺癌患者中的表达水平与患者的性别、年龄、病理类型、淋巴结转移情况和TNM分期等因素无关（均P〉0．05）。两种自身抗体联合检测的诊断敏感度为63．0％，特异度为91．2％，ROC曲线下面积为0．881，诊断效能优于单一自身抗体。结论血清p53、Bmi-1自身抗体联合检测具有较高的诊断价值，可能有助于肺癌的辅助诊断。

抗角蛋白抗体与类风湿因子检测在老年起病类风湿关节炎中诊断价值的分析

目的研究抗角蛋白抗体（抗AKA抗体）和类风湿因子（RF）的检测在老年起病的类风湿关节炎（EORA）诊疗中的临床价值。方法分别用间接免疫荧光法、BeckMan全自动蛋白分析仪同时检测EORA组35例，非老年起病的类风湿关节炎（NEORA）75例，16例正常老年人血清抗AKA抗体和RF。结果抗AKA抗体和RF对EORA诊断的灵敏度分别为57．1％、71．4％，特异性分别为100％、68．8％；对NEORA诊断的灵敏度分别为36％、88％。抗AKA抗体在EORA组的敏感性高于NEORA组，而RF在NEORA组的敏感性则高于EORA组（P〈0．05），在EORA组中AKA的特异性高于RF。结论联合检测抗AKA抗体、RF有助于提高老年起病类风湿关节炎的诊断率。

血清抗核周因子抗体(APF)在类风湿关节炎诊断中的应用

目的:探讨杭核周因子抗体(APF)对于早期类风湿关节炎(RA)的诊断价值。方法:对70例早期RA患者、90例非RA性自身免疫性疾病患者和50例正常对照者的血清APF和RF进行了测定比较。结果:RA组中APF阳性55例,RF阳性50例;非RA组中无APF阳性,RF阳性25例;正常对照组中无APF阳性,RF阳性1例。APF的特异性为100.0%,RF的特异性为81.4%。经统计学处理,RF的特异性与APF的特异性相比有统计学差异(P<0.01)。结论:APF是RA的特异性血清指标,对早期RA的诊断具有一定价值。

抗中性粒细胞胞浆抗体及抗酿酒酵母抗体在溃疡性结肠炎诊断中的应用

目的探讨抗中性粒细胞胞浆抗体(pANCA)、抗酿酒酵母抗体(ASCA)在溃疡性结肠炎(UC)诊断、鉴别诊断中的意义及pANCA与UC活动性的关系。方法采用间接免疫荧光法测定血清pANCA和ASCA水平。结果 pANCA诊断UC的敏感性、特异性分别为62.5%、96.1%,联合监测pANCA+/ASCA-诊断UC的敏感性、特异性分别为52.5%、100%。pANCA阳性率高低与UC活动状态、严重程度、病变范围无关。结论 pAN-CA是UC较特异血清抗体,可用于鉴别UC和克罗恩病(CD)。联合监测pANCA/ASCA可进一步提高诊断UC的特异性。但pANCA与UC活动性、病情轻重程度、病变范围无关。

自身抗体在流产诊断中的应用

目的:探讨甲状腺自身抗体、抗卵巢抗体(AoAb)对流产的影响,进一步揭示流产的病因。方法:用ELISE法和化学发光法对80例流产妇女血清AoAb和甲状腺球蛋白抗体(a-TG)、甲状腺过氧化物酶抗体(TPOAb)进行检测,并与40例正常生育妇女血清作对照。结果:流产组a-TPO阳性率为25.0%,高于正常对照组,统计学上有差异。a-TG的阳性率在RSA妇女中18.8%,高于正常对照组,有统计学意义。流产组AoAb阳性率为13.48%,显著高于对照组。结论:a-TG与a-TPO和AoAb均与流产有密切关系。

Application of high-titred IgY antibodies in orthopox virus diagnostics

Layer chickens were immunized with three species of inactivated orthopox virus （vaccinia virus, calpox virus and cowpox virus）. Antibodies （IgY） were purified from egg yolks by improved polyethylene glycol precipitation. The development of IgY directed against orthopox virus antigens was followed by immunofluorescence assay, plaque reduction neutraliztion test and immunoelectron microscopy. Cross-reactivity of two IgY antibodies with cells infected by the different strains of the pox viruses was also investigated by different methods （immunofluorescence assay, plaque reduction neutraliztion test and Western blot）. Even in very high dilutions in immunofluorescence assay （titres up to 1：10^6 and 1：10^5, respectively） and persisted on a plateau over 10 months after four booster injections, it was showed that anti-vaccinia virus IgY and anti-calpox virus IgY were positive. Neutralizing activity and ultra-structural detection of antigen with gold-labelled antibodies were respectively observed in plaque reduction neutralization test and immunoelectron microscopy. Western blot analysis revealed specific binding of IgY to virus proteins. Thus, there was cross-reactivity between different orthopox viruses. Finally, orthopox virns-specific IgY antibodies bounded magnetic beads （Dynabead） were used to concentration of orthopox viruses. This study suggests that anti-pox virus IgY could serve as a useful tool for orthopox viruses diagnosis.

Diagnosis of intestinal tuberculosis using a monoclonal antibody to Mycobacterium tuberculosis

AIM:To investigate the utility of immunohistochemical(IHC) staining with an antibody to Mycobacterium tuberculosis(M.tuberculosis) for the diagnosis of intestinal tuberculosis(TB).METHODS:We retrospectively identified 10 patients(4 males and 6 females;mean age = 65.1 ± 13.6 years) with intestinal TB.Clinical characteristics,including age,gender,underlying disease,and symptoms were obtained.Chest radiograph and laboratory tests,including sputum Ziehl-Neelsen(ZN) staining,M.tuberculosis culture,and sputum polymerase chain reaction(PCR) for tubercle bacilli DNA,as well as Tuberculin skin test(TST) and QuantiFERON-TB gold test(QFT),were examined.Colonoscopic records recorded on the basis of Sato's classification were also reviewed,in addition to data from intestinal biopsies examined for histopathological findings,including hematoxylin and eosin staining,and ZN staining,as well as M.tuberculosis culture,and PCR for tubercle bacilli DNA.For the present study,archived formalin-fixed paraffin-embedded(FFPE) intestinal tissue samples were immunohistochemically stained using a commercially available species-specific monoclonal antibody to the 38-kDa antigen of the M.tuberculosis complex.These sections were also stained with the pan-macrophage marker CD68 antibody.RESULTS:From the clinical data,we found that no patients were immunocompromised,and that the main symptoms were diarrhea and weight loss.Three patients displayed active pulmonary TB,six patients(60%) had a positive TST,and 4 patients(40%) had a positive QFT.Colonoscopic findings revealed that all patients had type 1 findings(linear ulcers in a circumferential arrangement or linear ulcers arranged circumferentially with mucosa showing multiple nodules),all of which were located in the right hemicolon and/or terminal ileum.Seven patients(70%) had concomitant healed lesions in the ileocecal area.No acid-fast bacilli were detected with ZN staining of the intestinal tissue samples,and both M.tuberculosis culture and PCR for tubercle bacilli DNA were negative in all samples.The histopathological data revealed that tuberculous granulomas were present in 4 cases(40%).IHC staining in archived FFPE samples with anti-M.tuberculosis monoclonal antibody revealed positive findings in 4 patients(40%);the same patients in which granulomas were detected by hematoxylin and eosin staining.M.tuberculosis antigens were found to be mostly intracellular,granular in pattern,and primarily located in the CD68 + macrophages of the granulomas.CONCLUSION:IHC staining with a monoclonal antibody to M.tuberculosis may be an efficient and simple diagnostic tool in addition to classic examination methods for the diagnosis of intestinal TB.

CLINICAL EVALUATION OF FOUR RECOMBINANT TREPONEMA PALLIDUM ANTIGEN-BASED RAPID TESTS IN THE DIAGNOSIS OF SYPHILIS

Objective To assess the sensitivity, specificity, and feasibility of 4 recombinant Treponema pallidum antigenbased rapid tests in the diagnosis of syphilis. Methods A total of 970 outpatients were selected from the Sexually Transmitted Diseases Centre of Peking Union Medical College Hospital. Venous blood was collected and serum was extracted. T. paUidum antibodies in whole blood, anticoagulant whole blood, and serum were detected using 4 recombinant T. pallidum antigen-based rapid tests. T. pallidum haemagglutination test （TPHA） was considered as the gold standard for the detection of T. pallidum specific antibodies in serum. The sensitivities and specificities of four methods were analyzed. Results The sensitivities and specificities of Abbott Determine Syphilis TP test, SD-BIOLINE Syphilis 3.0 test, VISITECT-SYPHILIS test, and Syphicheck-WB test for serum specimens were 100% and 98. 9%, 95.7% and 98.0%, 94.6% and 98.2%, 68.1% and 98.9% ; for whole blood were 74. 1% and 99. 5%, 87.9% and 99.4% , 73.2% and 99.7%, 64. 7% and 99.7%. The observed sensitivities of the 4 rapid diagnosis tests were not significantly different with TPHA （ P 〉 0.05 ）. Conclusions The 4 rapid tests show good performance and characteristics in the diagnosis of syphilis. Furthermore, they are more sensitive for serum specimens than whole blood.

The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

Development of a Polyclonal Antibody-based AC-ELISA and Its Comparison with PCR for Diagnosis of Canine Parvovirus Infection

A polyclonal antibody-based antigen-capture ELISA （AC-ELISA） has been developed for detection of Canine parvovirus （CPV） antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1：1 600 and 1：400, respectively, in the check-board titration. In this study, a total of 152 samples （129 faecal samples and 23 cell culture supernatant） were tested both by AC-ELISA and by polymerase chain reaction （PCR）. Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 102.8 TCID50/mL whereas PCR sensitivity was 100.8 TCIDs0/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.

^(99)Tc^m direct labeling of angiostatin

Objective: To explore the method of 99 Tc m direct labeling of angiostatin (AS) and investigate the stability and bioactivity of the 99 Tc m labeled AS in vitro . Methods: AS was extracted, validated, and then labeled with 99 Tc m after having been reduced by 2 ME or SnCl 2. The best labeling condition was screened by cross design. The labeling efficiency was measured by TLC and column chromatography. The stability of 99 Tc m AS was observed and compared when BSA, saline and different molar ratios of Cys∶AS were separately added. The bioactivity of 99 Tc m AS was observed in human umbilical vein endothelial cell (CEV304). Results: The labeling efficiency can reach (97±1 5)% for the 2 ME reducing approach. Its best experimental condition was as follows: AS 100 μg,PB(0 5 mol/L, pH 7 3)1 ml, 2 ME 100 μg, MDP (dissolved in 1 ml saline) 10 μl, and 99 Tc mO 4 - 185 MBq. The labeling efficiency using SnCl 2 reducing method can reach (90±3 0)%. The best experimental procedure was as follows: AS 100 μg,boric acid buffer(0 1 mol/L, pH 9 0)1 ml, 2%SnCl 2 (dissolved in 1 mol/L hydrochloric acid) 20 μl, was added into MDP, which was diluted with 1 ml deoxygenized water, and then 20 μl, 99 Tc mO 4 - 185 MBq was added. The product of 99 Tc m labeled AS was stable in vitro and had the same bioactivity as AS. Conclusion: 99 Tc m direct labeling of AS is simple and efficient. And the bioactivity of 99 Tc m AS has no significant change compared with AS.

Science Letters:Demonstration of a new biosensing concept for immunodiagnostic applications based on change in surface conductance of antibodies after biomolecular interactions

We report an important observation that the surface conductivity of antibody layer immobilized on polylysine-coated glass substrate decreases upon the formation of complex with their specific antigens. This change in conductivity has been observed for both monoclonal and polyclonal antibodies. The conductance of monoclonal mouse 1gG immobilized on polylysine-coated glass substrate changed from 1.02×10^-8 Ω^-1 to 1.41×10^-11 Ω^-1 at 10 V when complex is formed due to the specific biomolecular interactions with rabbit anti-mouse 1gG F（ab′）2. Similar behavior was observed when the same set up was tested in two clinical assays： （1） anti-Leishmania antigen polyclonal antibodies taken from Kala Azar positive patient serum interacting with Leishmania promastigote antigen, and （2） anti-p21 polyclonal antibodies interacting with p21 antigen. The proposed concept can represent a new immunodiagnostic technique and may have wide ranging applications in biosensors and nanobiotechnology too.

Development of Co-agglutination Method for Viral Nervous Necrosis from Grouper （Epinephelus sp,） in Batam

Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, rapid and reliable diagnostic test suitable for use in the field or laboratory without any special apparatus. The study aimed to study application of a co-agglutination test using Staphylococci sensitized with specific antibody for the diagnostic of VNN in grouper. First, brain and eye organ samples from diseased fish are homogenized with phosphate buffer saline （PBS） of pH 7.2. Then, the supernatant is collected after centrifugation at 8,000 rpm for 20 min, and finally one drop of the supematant and one drop of anti VNN antibody sensitized Staphylococci suspension are mixed on a glass slid and observation of the agglutination is performed after 5-10 min. The result shows that co-agglutination technique detects positive VNN in the brain and eye samples. The co-agglutination technique may provide valid result in a very short time as compared with complex method, such as enzyme-linked immunosorbant essay （ELISA） and fluorescient antibody technique （FAT） that require a high cost. Thus, this test can detect VNN faster, simple and more economic.