微波酶联免疫吸附试验诊断日本血吸虫病

目的评价微波酶联免疫吸附试验(微波ELISA)诊断日本血吸虫病的效果.方法用微波ELISA和常规ELISA平行检测日本血吸虫病各期患者血清262份,健康人血清92份,华支睾吸虫和卫氏并殖吸虫患者血清各15份.结果两种方法的日本血吸虫病患者血清抗体阳性检出率分别为89.69%(235/262)和91.98%(241/262),差异无显著性.健康人血清抗体假阳性率分别为2.17%(2/92)和1.09%(1/92),差异亦无显著性.均发现1例华支睾吸虫病患者的交叉反应.结论微波ELISA具有较高的敏感性和特异性,且具有快速获得检测结果的特点.     

血吸虫循环抗原检测及疗效考核价值

采用单克隆抗体(McAb)S31/32双夹心ELISA法检测感染不同寄生虫病人血清中循环抗原(CAg)427份,其中慢性血吸虫病188例,阳性检出率为95.2%,晚期血吸虫病阳性率4.5%(2/44),正常53人假阳性率5.7%.测定其他寄生虫感染患者及肝炎病人,除发现肺吸虫病、旋毛虫病和华枝睾吸虫病有交叉反应外,其它均呈阴性反应.同时,检测血吸虫病治疗前、后CAg,计495 份血清,慢性血吸虫病治疗前188例,阳性率95.2%,吡喹酮治疗后3、6、12和36个月,其阴转率分别为20.4%、31.9%、96.2%和87.7%.比较双夹心ELISA法和一步法ELISA检查CAg,无显著差异.测试结果提示,采用单抗S31/32建立的双夹心ELISA检测血吸虫循环抗原有很高的敏感性和良好的特异性,病人在治疗后一年其阴转率高,有很好的疗效考核价值.     

IEST、IFAT和Dot-ELISA诊断华枝睾吸虫病的比较研究

用成虫冰冻切片抗原免疫酶染色试验(IEST)、间接免疫荧光抗体试验(IFAT)和斑点-ELISA(Dot-ELISA)对比检测了51例华枝睾吸虫病人血清,阳性率分别为92%、88%和92%.检测健康人血清50例,假阳性率分别为2%、4%和2%.用3法检测22例急性血吸虫病人血清、20例慢性血吸虫病人血清和15例肺吸虫感染者血清.IEST的交叉反应率分别为14%、5%和0;IFAT的交叉反应率分别为14%、10%和0;Dot-ELISA的交叉反应率分别为14%、5%和0.显示3种方法诊断华枝睾吸虫病均有较高的敏感性和特异性.而IEST和Dot-ELISA更适于现场应用.     

我国家禽戊型肝炎病毒感染情况的调查

目的了解我国家禽戊型肝炎病毒(HEV)感染情况。方法自我国15个省份采集鸡和鸭血清共计1647份,应用酶联免疫吸附试验(ELISA)分别检测血清中HEV抗原和抗体;通过Nested PCR对陕西、西藏和海南的全部血清样本及其它抗原阳性样本分别进行HEV及禽HEV核酸检测。结果 1507份鸡血清中有54份为HEV抗体阳性,26份为HEV抗原阳性,抗原和抗体的阳性率分别为3.58%和1.73%;140份鸭血清中10份为HEV抗体阳性,2份为HEV抗原阳性,阳性率分别为7.14%和1.43%;鸡和鸭HEV抗原、抗体阳性率无统计学差异。在鸡、鸭血清标本中没有检测到HEVRNA。结论我国家禽存在HEV感染,但还需要进一步研究以确定感染的病毒是否为禽HEV或1-4型HEV跨种感染。     

双抗体夹心酶联免疫吸附试验(S-ELISA)检测血吸虫病人循环抗原的研究

应用从重感染兔血清中提取的γ-球蛋白为捕捉抗体、以酶标单克隆抗体为结合物进行双抗体夹心酶联免疫吸附试验(S-ELISA),检测日本血吸虫病人循环抗原.结果:69例急性血吸虫病人血清的阳性率为80.6%一90.9%;110例慢性血吸虫病人血清的阳性率为88.2%;40例华支睾吸虫病人血清有1例出现阳性,交叉反应率为2.5%,50例健康人血清亦有1例出现阳性反应,假阳性率为2%.提示该法具有较高的敏感性、特异性,判断结果较客观,可应用于现场.     

PVF抗原片环卵沉淀试验诊断日本血吸虫病敏感性和特异性的进一步观察

据80份粪检阳性的血吸虫病人血清,95份经吡喹酮治疗后3～8年的血吸虫病患者血清和90份非流行区健康人血清,按单盲法混合编号的考核实验结果:PVF—COPT使用江苏省寄研所提供的冷冻干卵抗原和热处理超声干卵抗原,对血吸虫病人血清的阳性率均为83.8％;在有血吸虫病治疗史者血清的阳性率分别为22.1％和15.8％;对正常人血清的假阳性率分别为8.9％和6.7％.此外,对肺吸虫、华枝睾吸虫病和姜片虫病人血清采用冷冻干卵抗原的交叉反应阳性率分别为8％、12％和4％。与常规法COPT的平行对比实验结果均无显著的差异.但标准化的PVF—COPT能预制定量的干卵抗原片,有利于提高COPT的稳定性.操作上较常规法COPT简便.     

重组Sj-Ts4样蛋白在日本血吸虫病免疫诊断中的初步应用

目的 评价重组Sj-Ts4样蛋白对日本血吸虫病的诊断价值.方法 取血吸虫病患者(急性、慢性、晚期)血清74份、华支睾吸虫病患者血清24份、钩虫病患者血清8份和健康人血清30份,利用纯化、重组rSj-Ts4样蛋白和日本血吸虫成虫抗原(SjAWA),采用ELISA法分别检测血清中抗,Sj-Ts4样蛋白特异性抗体和SjAWA抗体,比较两种检测方法的敏感性和特异性.结果 rSj-Ts4-ELISA和SjAWA-ELISA法检测急性、慢性和晚期血吸虫病患者血清的阳性率分别为97.1%(33/34)、100.0%(16/16)、87.5%(21/24)和100.0%(34/34)、100.0%(16/16)、75.0%(18/24),经校正X2检验,两种检查方法比较差异无统计学意义(x2=1.23.P＞0.05);健康人血清两种检测方法的假阳性率分别为6.7%(2/30) 3.3%(1/30),经校正x2检验,差异无统计学意义(X2=0.35,P＞0.05);华支睾吸虫患者和钩虫病患者血清阳性率分别为12.5%(3/24)、20.8%(5/24)和12.5%(1/8)、37.5%(3/8),经校正x2检验,两种检查方法比较差异无统计学意义(x2值分别为0.60、1.33,P＞0.05).结论 rSj-Ts4样蛋白作为抗原,对免疫检测日本血吸虫病患者血清中的抗体具有较好的敏感性和特异性,对日本血吸虫病患者的诊断有较大的应用价值.     

不同血清学方法对调查现场包虫病人血清的检测及评价

本文应用青海囊型包虫抗原 Dot- EL ISA、IHA和新疆囊型包虫抗原 EL ISA、泡型包虫 EM18抗原 EL IB对流行病学调查现场的 2 0 6例包虫病人血清进行了检测。结果表明青海囊型包虫抗原 Dot- EL ISA和 IHA对囊型包虫病人血清的阳性率分别是 90 .37%和 91.98% ;新疆囊型包虫抗原 EL ISA对囊型包虫病人血清的阳性率为 75 .94% ;三种方法对钙化灶型囊型包虫病人血清的阳性率分别是 77.2 7%、81.82 %和 6 5 .91% ,显著低于其它囊型包虫病人血清的阳性率 ;阴性者主要是单纯性肺脏和肝脏囊型包虫病人血清。3种方法 B超、X线诊断的泡型包虫病人的血清阳性率均为 10 0 .0 %。泡型包虫抗原 EM18-EL IB检测结果表明 ,B超 ,X线诊断的泡型包虫病人血清阳性率为 73.6 8% ;囊型包虫病人血清阳性率为 5 .88% ,其中钙化灶型病人血清阳性率为 15 .91% ;阳性血清数与阴性血清数比例约为 1:7(2 5 :181)。对不同方法在囊型和泡型包虫病诊断与鉴别诊断中的价值及意义进行了讨论     

滴金免疫测定法用于检测囊虫病患者循环抗原的研究

目的　建立一种敏感、快速、简便的囊虫病诊断方法。方法　应用两株抗猪囊尾蚴囊液抗原的单克隆抗体(McAb) ,一株滴于硝酸纤维素膜上 ,另一株与胶体金结合为探针 ,建立了检测囊虫病患者血清中循环抗原 (CA)的滴金免疫测定法 ,抗原、抗体通过渗滤在膜上进行反应 ,10min内即可用肉眼观察结果。对不同稀释度的猪囊尾蚴囊液抗原进行检测 ,确定本法的最低抗原检出量 (0 .4 5ng/ml)。将本法与ELISA作了比较。 结果　对 78例囊虫病患者血清进行检测 ,循环抗原阳性率为 87.18%。其中活动型脑囊虫病患者 5 2例 ,循环抗原阳性率为 98.0 8% ;非活动型脑囊虫病患者 2 0例 ,循环抗原阳性率为 6 5 .0 0 % ;单纯皮肌型囊虫病患者 6例 ,循环抗原阳性率为 6 6 .6 7%。 4 0份健康人血清仅有 1例 (2 .5 0 % )出现假阳性 ,与10例华支睾吸虫病患者血清、15例血吸虫病患者血清及 2 3例非囊虫病的脑部疾病患者血清均无交叉反应。该方法检出最低猪囊尾蚴囊液抗原量为 0 .4 5ng/ml。本法与ELISA的符合率为 98.19%。结论　DIGFA简便、快速 ,结果客观 ,对囊虫病特别是活动型脑囊虫病敏感性高 ,特异性强 ,值得进一步推广应用。     

ELISA双抗原夹心法检测内脏利什曼病抗体的研究

目的观察rK39抗原酶联免疫吸附试验双抗原夹心法(ELISA夹心法)检测内脏利什曼病抗体用于内脏利什曼病诊断与宿主动物感染调查的可行性。方法采用ELISA夹心法与rK39免疫层析试条法(ICT)同步检测内脏利什曼病抗体。结果 5种家畜血清229份,7种鼠类标本238份,ELISA夹心法和rK39 ICT法抗体检测均阴性;接种利什曼原虫灰仓鼠、草原兔尾鼠标本33份,13份阳性,阳性率39.4%;塔里木兔标本119份,阳性7份,阳性率5.9%;非疫区儿童血清29份,全部阴性;疫区无内脏利什曼病症状人血清250份,4份两种方法均阳性,阳性率均为1.6%;住院内脏利什曼病病人血清67份,两种方法的阳性率分别为68.7%和67.2%。共检测人和其他动物标本965份,两种方法的阳性符合率98.6%,阴性符合率99.9%。ICT相同显色等级的阳性标本,ELISA跨10个滴度。结论 ELISA夹心法可检测多种动物内脏利什曼病抗体,抗体滴度具有定量意义。该方法适用于内脏利什曼病诊断、疗效观察和流行病学调查。     

包虫病人血清特异性抗包虫IgG和IgM的检测及其临床意义探讨

本文报告应用IFAT法检测包虫病人血清中特异性包虫IgG和IgM的初步结果。44例正常人血清特异性IgG和IgM的假阳性率均为0％,83例包虫病患者血清特异性IgG和IgM的阳性检出率分别为96.36％和19.28％,二者有极显著性差异(P<0.01)。提示现症包虫病患者血清中的特异性抗包虫抗体主要为IgG。在16例特异性IgM阳性病人中,有9例存在包虫囊破裂,7例为肺包虫或肝肺混合包虫病患者,表明特异性IgM在包虫囊破裂时升高,肺包虫病人比肝包虫病人检出率高。提示包虫病人血清中的特异性IgM抗体水平与包虫囊的生活状态有关。     

Serological survey of normal humans for natural antibody to cell surface antigens of melanoma.

Sera of 106 normal adult men were tested for antibodies reacting with cell surface antigens of three established lines of cultured malignant melanoma. Positive reactions with a protein A assay for IgG antibodies were extremely rare (1-2%). The frequency of positive reactions with assays for IgM antibodies was higher: 5-15% in immune adherence assays and 55-82% in anti-C3 mixed hemadsorption assays. After low-titered sera and sera reacting with fetal calf serum components, conventional alloantigens, and widely distributed class 3 antigens were excluded, sera from seven individuals (one with IgG antibody and six with IgM antibodies) were selected for detailed analysis. The serum containing the IgG antibody came from a healthy 65-year-old Caucasian man; titers of antibody in his serum ranged from < 1/10 to 1/40,000 in tests with different melanoma cell lines. This IgG antibody identifies a differentiation antigen of melanocytes, provisionally designated Mel 1, that distinguishes two classes of melanomas: 22 melanoma cell lines typed Mel 1+ and 17 types Mel 1-. Mel 1 is expressed by fetal fibroblasts but not adult fibroblasts and can be found on a proportion of cultured epithelial cancer cell lines (5 out of 23) but not on glioma or B-cell lines. The melanoma antigens detected by the naturally occurring IgM antibodies are serologically unrelated to Mel 1 but, like Mel 1, appear to be differentiation antigens that distinguish subsets of melanoma. These IgM antibodies detect antigens that are identical or closely related to the AH antigen, a melanoma surface antigen that was initially defined by autologous antibody in a patient with melanoma. In view of the immunogenicity of both Mel 1 and the AH antigens in humans and their occurrence on more than 50% of melanomas, it remains to be seen whether antibody to these antigens can be elicited by specific vaccination of seronegative melanoma patients and whether this will have an influence on the clinical course of the disease.     

Recognition of Treponema pallidum antigens by IgM and IgG antibodies in congenitally infected newborns and their mothers

Immunoblotting techniques were used to examine the proteins of Treponema pallidum recognized by IgM and IgG antibodies in sera from infants with congenital syphilis and their mothers. Infected infants' serum IgM reactivity to treponemal antigens differed from that of control infants born to normal, serofast, and biologic false-positive mothers. Each of the infected infants' sera exhibited IgM reactions to the 47- and 37-kilodalton (kDa) proteins of T. pallidum. Although rheumatoid factor was detected in the sera of half of the infected infants, removing this factor did not alter the pattern of IgM blots. IgG reactions in infants were almost exclusively of the IgG1 and IgG3 subclasses and mirrored those of the mother, except for IgG1 and IgG3 reactions to the 83-kDa treponemal protein, which were unique to infants' sera. Our results suggest that the findings of IgM antibody directed against the 47- or 37-kDa antigens of T. pallidum may help to diagnose congenital syphilis at birth.     

棘球蚴病患者IgG抗体阴性反应血清再检测的研究

目的　探索棘球蚴病患者抗体应答假阴性反应原因 ,以改进棘球蚴病的免疫诊断方法。　方法　采用间接ELISA和双抗体夹心ELISA方法 ,检测 42例IgG抗体阴性反应棘球蚴病患者血清的IgG亚类 (IgG1、IgG2、IgG3和IgG4 )、IgA、IgM、IgE抗体及抗原和循环免疫复合物。　 结果　 42例阴性血清中 ,32例IgG亚类或IgA、IgM、IgE抗体阳性 ,1 0例血清抗体全部阴性。其中IgG1、IgG4及IgA、IgM、IgE的检出率明显高于正常人 ,分别为 42 .9%、1 1 .9%、2 8.6 %、2 6 .2 %和 2 1 .4 %。小儿的IgM高于成人。肝棘球蚴病患者的IgG亚类高于肺棘球蚴病患者。IgG1与其它抗体联合检测 ,以IgG1 +IgA +IgM检出率最高 ,为 64 .3 %。IgG阴性患者血清的CAg和CIC阳性率分别为 2 8.57%及30 .95 %。　结论　抗棘球蚴总IgG抗体表达水平低下 ,抗体表达种类不同及循环免疫复合物的形成 ,是造成棘球蚴病患者IgG抗体反应阴性的主要原因。IgG1 +IgA +IgM检测可提高棘球蚴病患者的诊断率     

TORCH感染与不良妊娠结局的相关性分析

目的 目的 了解TORCH感染与不良妊娠结局间的关系。方法 方法 采用捕获ELISA法对900例孕妇血清进行TORCH? IgM抗体检测, 对阳性孕妇进行追踪, 并随访其妊娠结局。结果 结果 孕妇血清TORCH?IgM抗体总阳性率为4.11% （37/900）,其中巨细胞病毒、 单纯疱疹病毒、 风疹病毒和弓形虫特异性IgM抗体阳性率分别为2.00% （18/900）、 0.78% （7/900）、 0.44% （4/900） 和0.89% （8/900）。31例继续妊娠的孕妇中, 不良妊娠结局20例, 占64.52%。结论 结论 TORCH感染是引起不良妊娠结局的重要危险因素。孕期进行TORCH筛查并给予适当干预, 可减少不良妊娠的发生, 预防出生缺陷。     

Mac ELISA测定乙脑患者血清和脑脊液中特异性IgM的评价

用抗体捕捉ELISA法(Mac ELISA)检测临床疑似流行性乙型脑炎(下称乙脑)病人脑脊液88份,阳性51份,阳性率57.95％;血清91份,阳性56份,阳性率为61.53％,两者无显著性差异(X~2=0.238 P>0.05);包括取自同一患者的脑脊液和血清各72份,阳性率分别为61.11％和68.05％;而分析其中14例IgG呈4倍以上增高,乙脑患者脑脊液IgM阳性13例,而血清IgM阳性却只有8例,提示在疫区流行季节内仅仅血清学IgM阳性有时可能为乙脑隐性感染的同时还伴其它有关病毒感染。     

Serological cross-reaction among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody method]

We studied the serological cross-reactions among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody (IFA) method, using sera from 8 patients with cat scratch disease (CSD), 13 patients with C. pneumoniae infection and 12 patients with acute Q fever. B. henselae IgG antibody was negative in 13 patients with C. pneumoniae infection, and was positive in 3 (titers being 1:64) of 12 patients with Q fever, whereas B. henselae IgM antibody was negative in all the patients with C. pneumoniae infection or Q fever. C. burnetii IgG antibody was removed by absorption of these 3 sera with C. burnetii antigens, whereas B. henselae IgG antibody did not change. C. pneumoniae IgG antibody was positive in 3 (titers being 1:125 in two, 1:32 in one) of 8 patients with CSD. Both C. pneumoniae and B. henselae IgG antibody titers were significantly reduced by absorption of these 3 sera with B. henselae antigens. C. burnetii IgG or IgM antibodies were negative in all patients with CSD. In conclusion, no serological cross-reaction between B. henselae and C. burnetii was observed. On the other hand. B. henselae IgG antibody cross-reacted to C. pneumoniae antigens, whereas C. pneumoniae IgG antibody did not cross-react to B. henselae antigens. Our findings suggest that determination of B. henselae IgG or IgM antibodies were not influenced by C. pneumoniae and C. burnetii antigens.     

Antibody response to a polysaccharide antigen present in the schistosome gut. I. Sensitivity and specificity.

By using paraffin sections of adult schistosomes fixed in Rossman's fixative, specific human IgM and IgG antibodies to a polysaccharide present in the epithelial cells of the schistosome were measured by using indirect immunofluorescent techniques. Of the 49 patients, mostly infected with S. mansoni but a few infected with S. haematobium or S. japonicum, all had antibody present at a 1:8 dilution of serum. Specific IgM antibody was more sensitive than IgG, yielding 100% and 86% positivity respectively. The false positive rate was 3% in a panel of sera obtained from patients most of whom were infected with other parasites. Antibodies were detected by the 3rd week in experimentally infected animals. Unisexual infections also induced antibody production. As a diagnostic test, the measurement of antibody to the polysaccharide is an easily performed reliable test with high sensitivity and specificity.     

Isotypic analysis, antigen specificity, and inhibitory function of maternally transmitted Plasmodium falciparum-specific antibodies in Gabonese newborns.

An analysis of Plasmodium falciparum-specific antibodies was performed in pairs of maternal and cord sera from Gabon, a region endemic for malaria. All paired sera (n = 59) had P. falciparum-specific antibodies. Immunofluorescence assays detected parasite-specific IgG1, IgG2, and IgG3 in 100% of the tested pairs (n = 26) and IgG4 in 42% of them. The titers of specific IgG2 and IgG3 were significantly lower in cord than in maternal sera. All maternal sera had specific IgM. Of the seven P. falciparum-IgM positive cord sera, six were associated with malaria-related histological placental changes (MRHPC). In addition, higher titers of specific IgG1 in maternal and cord sera and of specific IgG3 in cord sera were associated with MRHPC. Similar P. falciparum antigens were recognized by cord and corresponding maternal sera in radioimmunoprecipitation and Western blot assays (n = 40). Sixteen of 20 cord sera and 15 of 20 paired maternal sera significantly inhibited in vitro parasite growth. The extent of inhibition did not correlate with the titer of specific antibodies. These data confirm the very effective placental transfer of anti-malarial antibodies. The presence of IgM in some cord sera raise the question of intrauterine sensitization to malaria antigens.     

抗旋毛虫肌肉期幼虫排泄分泌物中46-58kDa蛋白IgM和IgG抗体的研究

目的寻求简便、快速、可靠的方法,用于旋毛虫病的诊断及疗效考核。方法建立检测抗旋毛虫肌肉期幼虫排泄分泌物(TsL1-ES)中46-58kDa蛋白抗体的斑点金免疫渗滤试验(DIGFA),并对患者在治疗前和治疗后血清中特异抗体水平的动态变化进行研究。结果旋毛虫病患者治疗前血清中特异IgM和IgG抗体的检出率分别为94.83%(55/58)和91.38%(53/58);正常人血清(100份)、囊虫病(25份)、血吸虫病(20份)、肺吸虫病(20份)及蛔虫病(22份)病人血清均未检出该两类特异性抗体。52例两类抗体均阳性的旋毛虫病患者经有效药物治疗后3个月、6个月、1年、1.5年连续采血,经DIGFA检测连续观察的结果表明,IgM抗体出现较早,阴转较为明显和迅速,治疗后3月的阴转率达46.15%(24/52),1年的阴转率可达88.46%(46/52);而IgG抗体则出现较晚,阴转缓慢,治疗后1.5年的阴转率仅为23.08%(12/52)。结论DIGFA为检测抗TsL1-ES中46-58kDa蛋白抗体的有效方法,检测IgM抗体可用于诊断及疗效考核,而IgG抗体的检测不宜用来判断疗效。