包虫病病人唾液中特异性抗体的研究

目的 :研究包虫病病人唾液中的特异性抗体的诊断价值。方法 :用 SPA夹心 EL ISA测定包虫病病人和正常人唾液中 Ig G总量。用抗原中和试验鉴定唾液 Ig G的特异性。结果 :32例包虫病病人和 32例正常人唾液中 Ig G总量分别为 10 5.6± 65.4 μg/ ml和 94 .2± 55.5μg/ ml,差异极显著 ( P<0 .0 1)。抗原中和试验 4 0例包虫病病人唾液 Ig G抗体反应抑制率为 30 %— 82 .1% ,32例正常人唾液无抑制效应。表明唾液中 Ig G抗体为特异性 Ig G抗体。对 89例包虫病病人和 12 9例正常人血清和唾液 Ig G- EL ISA检测结果 ,病人血清阳性率为 92 .1% ,唾液为 78.7%。唾液与血清的阳性符合率为 80 .5% ,阴性符合率为 98.3%。结论 :在包虫病病人唾液中测出特异性 Ig G抗体 ,可作为包虫病的辅助诊断。     

ABC-ELISA法诊断肠螨病的研究

目的　探讨生物素 -亲和素酶联免疫吸附试验 (ABC -ELISA)在肠螨病诊断中的应用价值。方法　采集 4 8例肠螨病患者血清 ,用ABC -ELISA法检测血清中螨特异性抗体IgG ,并与葡萄球菌A -蛋白酶联免疫吸附试验 (SPA -ELISA)进行比较。结果　用ABC -ELISA法和SPA -ELISA法检测 4 8例肠螨病患者血清中的螨特异性抗体IgG阳性率分别为89 5 8% (43/ 4 8)和 5 6 2 5 % (2 7/ 4 8) ,两者比较 ,差异具显著性 (χ2 =13 5 0 ,P <0 0 1)。结论　ABC -ELISA法是实验室诊断肠螨病的一种有效方法     

四川地区血吸虫及其他寄生虫病人血清库保存血清的质量探讨

目的　探讨四川地区血吸虫及其他寄生虫病人血清库的血清保存时间和效果。方法　分别于 1 994年、1 999年、2 0 0 3年抽取血清库内同一批慢性血吸虫病人和其它寄生虫病人血清以及非流行区的健康人血清 ,采用一步法 EIA-kit检测血吸虫病人血清循环抗原 ,用快速 ELISA、IHA检测血吸虫抗体。结果　三年测定血吸虫病人血清 EIA-kit阳性率分别为 1 0 0 .0 0 %、96.2 5 %、97.96% ,健康人血清的特异性均为 1 0 0 .0 0 %。 ELISA的阳性率分别为 1 0 0 .0 0 %、97.5 0 %、98.47% ;健康人血清的特异性为 99.44%、99.69%、99.62 % ;IHA的阳性率分别为 99.70 %、97.92 %、96.43 % ,健康人血清特异性为 98.61 %、98.44%、98.48%。用 ELISA对 3 69份其他寄生虫病人血清 (华支睾吸虫病 ,肺吸虫病 ,包虫病 ,囊尾蚴病 ,旋毛虫病 )检测抗体的特异性分别为 1 0 0 .0 0 %( 3 69/3 69)、97.67% ( 2 5 2 /2 5 8)、96.40 % ( 1 0 7/1 1 1 )。三次检测结果差异均无显著性 ( P>0 .0 5 )。结论　四川地区血吸虫及其它寄生虫病人血清库血清循环抗原、抗体效果 1 0年内无显著降低 ,有长期的应用价值     

旋毛虫成虫排泄分泌抗原检测唾液中抗旋毛虫IgG抗体的研究

目的 评价旋毛虫(Trichinella spiralis)成虫排泄分泌抗原(adult worm excretory-secretory antigen,AWESA)作为诊断抗原检测旋毛虫感染日本大耳兔唾液中抗旋毛虫IgG抗体的可行性. 方法 建立旋毛虫感染日本大耳兔和对照组兔动物模型,采集感染前和感染后1～6周兔唾液和血清以及对照组兔唾液和血清.制备AWESA,建立AWESA作为诊断抗原的间接酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA),以市售旋毛虫IgG抗体检测试剂盒作为对照,测定感染前和感染后1～6周兔唾液和血清以及对照组兔唾液和血清中抗旋毛虫IgG抗体.AWESA和试剂盒测得的唾液A值和血清A值进行线性相关分析,AWESA和试剂盒测得的唾液阳性率和血清阳性率分别进行x2检验.结果 AWESA检测唾液和血清特异性IgG抗体阳性率依次为0、5%、20%、40%、60%、85%、90%,0、30%、60%、85%、95%、100%、100%,除感染后0、1、2周外其余各周唾液A值与血清A值呈显著线性相关(P＞0.05、P＞0.05、P＞0.05、P＜0.05、P＜0.05、P＜0.05、P＜0.05).市售旋毛虫IgG抗体检测试剂盒检测旋毛虫感染前和感染后兔唾液和血清中特异性IgG抗体阳性率依次为0、15%、20%、40%、55%、75%、90%,0、35%、60%、95%、95%、100%、100%,除0、1、3周外其余各周唾液A值与血清A值呈线性相关(P＞0.05、P＞0.05、P＜0.05、P＞0.05、P＜0.05、P＜0.05、P＜0.05). 结论 AWESA与市售试剂盒检测唾液和血清中抗旋毛虫IgG抗体的阳性率具有一致性.     

胶体金免疫层析试纸条对云南鼠疫现场材料的检测

目的评价检测鼠疫抗原及抗体胶体金免疫层析试纸条(G ICA及RG ICA)对现场材料的检测效果。方法采用G ICA分别对采自云南省的607份血清标本(查鼠疫抗体)及572份鼠脏器标本(查鼠疫抗原)进行检测,同时以血凝试验(IHA及R IHA)与酶联试验(ELISA及F1-ELISA)作为对照。结果(1)在对607份血清标本的检测中,G ICA、IHA及ELISA三种方法的符合率中度,而G ICA的敏感性比IHA与ELISA分别高111%和90%;(2)在对572份鼠脏器标本的检测中,RG ICA、R IHA及F1-ELISA三种方法的符合率中度,而RG ICA的敏感性比R IHA与F1-ELISA分别高100%和14.3%。结论检测鼠疫的G ICA及RG ICA特异性强、灵敏度高、简便快速,有较大的推广应用价值。     

检测唾液中特异性抗体诊断人体旋毛虫病的研究

目的探讨检测唾液特异性抗体对人体旋毛虫病的诊断价值。方法用ELISA测定57例旋毛虫病人唾液中旋毛虫IgG抗体,并与血清同种抗体检测结果进行比较。结果检测唾液中旋毛虫IgG抗体诊断旋毛虫病的敏感性为64.91%,特异性为100%;检测血清旋毛虫IgG抗体诊断旋毛虫病的敏感性和特异性分别为91.23%和95.83%;唾液旋毛虫IgG与血清旋毛虫IgG的OD值呈正相关(r=0.4011)。结论检测唾液中特异性抗体对于诊断人体旋毛虫病有一定的应用价值,在采集血清有困难时可将唾液作为血清的替代检测标本。     

人唾液、尿液中弓形虫抗原抗体的检测

目的探讨用唾液、尿液替代血清进行弓形虫感染免疫学诊断的可行性。方法用ELISA法对血清进行弓形虫循环抗原(CAg)、IgM、IgG抗体的检测,取上述各项免疫学指标阳性的血清样本80例,并随机抽取3项指标均为阴性的血清样本102例,分别与其唾液和尿液标本在同一反应板上同步进行测定,比较其阳性和阴性符合率。结果(1)554份血清样本中,CAg、IgM和IgG3项的阳性率分别为4.51%(25/554)、3.61%(20/554)、6.32%(35/554)。(2)唾液中检出IgM抗体阳性16例,与血清的阳性符合率为80%(16/20),检测CAg、IgG抗体均为阴性。(3)检测尿液中的抗原抗体均无一例阳性。结论采用唾液标本检测弓形虫IgM抗体,在弓形虫感染的诊断及流行病学调查中均具有一定的应用前景。     

ELISA法检测血清包虫IgG抗体试剂盒的应用评价

目的对ELISA(酶联免疫吸附试验)法检测血清包虫IgG抗体试剂盒进行评价,为其应用提供参考。方法收集2012-01-12就诊的1242位可疑包虫病患者的血清标本,采用ELISA检测血清棘球蚴IgG抗体。结果1242例中就诊患者中对包虫感染有明确诊断(确诊或排除包虫病)的有1130例,其中棘球蚴IgG抗体阳性159例(14.07%),棘球蚴IgG抗体阴性971例(85.93%);以临床诊断为标准得到灵敏度87.06%,特异性91.87%,Youden指数为0.78。结论ELISA法检测棘球蚴IgG抗体的灵敏度和特异性较高,可用于包虫病患者的流行病学调查;并结合B超等影像学检查用于临床包虫病的辅助诊断。     

日本血吸虫重组表膜蛋白Tetraspanin 2免疫诊断价值初探

目的 探讨日本血吸虫重组表膜蛋白Tetraspanin 2(rSjTsp2)作为免疫诊断抗原的潜能.方法 利用表达纯化的rSjTsp2建立间接ELISA法(rSjTsp2-ELISA)检测不同感染时期的感染日本血吸虫家兔血清特异性IgG抗体,并与成虫粗抗原(AWA)-ELISA法进行比较.结果 rSjTsp2-ELISA对感染日本血吸虫2、4、6周家兔血清特异性IgG抗体检测的敏感性分别为75.4%(46/61)、77.6%(45/58)和81.5%(44/54),AWA-ELISA法敏感性分别为68.9%(42/61)、86.2%(50/58)和88.9%(48/54),同一时期两种方法检测抗体的效果比较,差异无统计学意义(x2值分别为0.652、1.454、1.174,P均＞0.05).结论体外成功表达了日本血吸虫表膜蛋白rSjTsp2,其原核表达重组蛋白具有一定的诊断应用潜能.     

ABC-ELISA法检测华支睾吸虫特异性IgG及其亚类

目的　研究ABC ELISA法检测IgG及IgG亚类用于华支睾吸虫病诊断的价值。　 方法　采用ABC ELISA法检测华支睾吸虫感染者 (62例 )和健康人 (3 8例 )血清特异性总IgG及其亚类。　结果　华支睾吸虫感染者血清特异性总IgG及其亚类阳性检出率分别为IgG 10 0 % (62 / 62 ) ,IgG15 4.8% (3 4/ 62 ) ,IgG2 79.0 % (4 9/ 62 ) ,IgG3 40 .3 % (2 5 /62 ) ,IgG498.4% (61/ 62 ) ;其中IgG4平均阳性滴度显著高于总IgG (P <0 .0 1)。健康人血清特异性总IgG假阳性率7.9% (3 / 3 8) ,IgG4假阳性率为 0 ;检测健康对照血清IgG4的A值 (0 .0 0 1)显著低于总IgG检测 (A =0 .0 3 1) (P <0 .0 1)。　结论　ABC ELISA法用于华支睾吸虫病诊断具有较高的敏感性 ;检测特异性IgG4诊断华支睾吸虫病效能与检测总IgG相当 ,同时具有阳性显色强、阴性本底低的优点 ;其余IgG亚类对于华支睾吸虫病诊断价值不大。     

Serodiagnosis of human paragonimiasis caused by Paragonimus heterotremus

Enzyme-linked immunosorbent assay (ELISA) and indirect hemagglutination tests (IHA) were evaluated for serodiagnosis of human paragonimiasis caused by Paragonimus heterotremus using homologous adult worm extract as antigen. IgG-ELISA was the most sensitive, being positive in all paragonimiasis sera tested while IHA and IgA-ELISA gave 88% and 59% positive rates respectively. Cross reactivity in IgG-ELISA was detected with fascioliasis sera, producing overall assay specificity of 97%. It is suggested that IgG-ELISA is a reliable serodiagnostic test for human paragonimiasis caused by Paragonimus heterotremus.     

同步检测血清和唾液中的棘球蚴抗原及其特异性抗体

应用ＭｃＡｂ－Ｓａｎｄｗｉｃｈ－ＥＬＩＳＡ（夹心法）及Ｄｏｔ－ＥＬＩＳＡ（点免疫法）同步检测了包虫病病人的不同抗原、抗体样品,结果表明ＭｃＡｂ夹心法对患血清、唾液抗原阳性检出率分别为８０．４３％和６０．８７％;点免疫法对血清、唾液抗体检出率为８６．９６％和７８．２６％,均明显高于对照组（Ｐ〈０．０５）,对抗原,抗体的同步检测结果进行显性及相关性分析,得出血清抗原检测可取代以往的血清抗体检测,唾     

酶联免疫吸附试验检测人血清弓形体IgG,IgM抗体的研究

对ELISA检测人血清弓形体IgG、IgM抗体进行了研究。450份孕妇血清中,ELISA阳性率显著高于IHA;抗体滴度分折,ELISA一般高于IHA2～10倍。25份ELISA IgG抗体阳性血清,IFA检出19份;3份IgM抗体阳性和2份IgG、IgM抗体均阳性血清,IFA分别检出2份。2份阴性和4份含不同抗体滴度的阳性血清于第一次测定后,第7天和第21天测定的阴、阳性结果一致,OD值变异系数为2.43～16.52％。39份阳性血清抗体滴度与OD值呈直线比例关系(r=0.991,P<0.0005)。结果表明,ELISA用于弓形体感染的血清学诊断具有较好的实用性。     

Comparison of serologic tests for the diagnosis and follow-up of alveolar hydatid disease

Alveolar hydatid disease is a serious and often fatal condition caused by infection with the metacestode form of Echinococcus multilocularis. Sera of 21 patients with histologically confirmed disease were tested by an enzyme-linked immunosorbent assay (ELISA) using a semi-purified E. multilocularis antigen fraction (Em2) and by indirect hemagglutination (IHA) and double diffusion (DD5) tests using antigens prepared from E. granulosus cyst fluid. At diagnosis, sera from all 21 patients were positive by Em2 ELISA, 18 (86%) by IHA, and 5 (24%) by DD5. Em2 ELISA detected an antibody response earlier than IHA in 4 of 9 patients from whom sera were available before diagnosis. Following complete surgical resection, Em2 ELISA converted from positive to negative in serum of 2 of 3 patients, while IHA results did not change. Following incomplete resection, 14 of 15 patients tested remained positive by Em2 ELISA, while 12 remained positive by IHA. Of sera from 361 healthy persons from regions free of E. multilocularis, none were positive by Em2 ELISA, while 8% were positive by IHA. Of sera from 59 patients with non-echinococcal parasitic infections, none were positive by Em2 ELISA, while 31% were positive by IHA. Thus, in comparison with tests using E. granulosus antigens, Em2 ELISA appears to be more sensitive and specific for diagnosing AHD, useful on follow-up of resected patients, and positive earlier in the course of disease.     

棘球蚴3种抗原在包虫病斑点酶联免疫吸附试验中特异性和敏感性比较

用羊源细粒棘球蚴囊液、原头蚴及生发层３种抗原,用酶标记羊抗人ＩｇＧ,对９４例包虫病患者、４４例非包虫患者及５０例健康人血清进行了斑点酶联免疫吸附试验。结果用囊液、原头蚴、生发层３种抗原的敏感性分别为９２．６％、９３．６％和９３．６％,５０例健康人血清均未出现假阳性反应,与各种肿瘤、各种炎症、各种肝病及结核患者血清有一定的交叉反应     

Enzyme-linked immunosorbent assay for the diagnosis of clinical and subclinical melioidosis

An enzyme-linked immunosorbent assay (ELISA) for the detection of specific IgG and IgM antibody to Pseudomonas pseudomallei was developed. The IgG-ELISA was compared with the indirect fluorescence assay for IgG antibody (IgG-IFA) and the indirect hemagglutination (IHA) test in studies with serum specimens from persons from endemic areas for melioidosis and from persons from nonendemic areas of Australia. The sensitivity and specificity of the IgG-ELISA were 90% and 99%, respectively, comparable to those obtained with the IgG-IFA. The IgG-ELISA was more sensitive than the IHA test (74%) and was more suitable than the IgG-IFA as a serologic screening test for melioidosis. The IgM-ELISA was compared with the IgM-IFA as a marker of disease stage in patients with melioidosis. There was good diagnostic agreement between the tests; 92% of patients with active disease gave IgM-ELISA titers greater than or equal to 1:5,120 and 93% of patients with subclinical melioidosis had IgM-ELISA titers less than or equal to 1:1,280. Of the overlap group of patients with a borderline IgM-ELISA titer of 1:2,560, approximately 33% were clinical cases. An uncommon disease stage consisting of a self-limited, short-term, flu-like, pyrexial illness accompanied by elevated serum IgM-ELISA titers (greater than or equal to 1:5,120) was seen in a small number of patients residing in endemic Australia.     

Dot－ELISA检测抗原与IHA、COPT检测抗体诊断血吸虫病的比较

本文比较了斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）检测循环抗原，间接血凝试验（ＩＨＡ）、环卵沉淀试验（ＣＯＰＴ）检测抗体诊断血吸虫病的效果。１２例急性血吸虫病患者血清３种方法检测均为阳性；２７７例慢性血吸虫病患者血清３种方法检测阳性率分别为７９．４２％、６４．６２％和６３．１８％。１４例华支睾吸虫病患者血清和８９例健康人血清Ｄｏｔ－ＥＬＩＳＡ检测均为阴性。１０例急性血吸虫病患者，在离开流行区经吡喹酮治疗后３７周时粪检全部转阴，检测血清循环抗原４例转阴，其余６例滴度大幅度下降；ＩＨＡ检测３例转阴，其余７例几何平均倒数滴度（ＧＭＲＴ）由４７．５７降至９．３５；ＣＯＰＴ检测平均环沉率由２１．５％降至４．１２％。结果表明Ｄｏｔ－ＥＬＩＳＡ检测血吸虫循环抗原具有较好的敏感性和特异性。这３种方法在血吸虫病诊断及疗效考核中均有一定价值。     

Detection of specific circulating antigen, immune complexes and antibodies in human hydatidosis from Turkana (Kenya) and Great Britain, by enzyme-immunoassay

Circulating antigen, specific immune complexes (IgG and IgM) and specific antibodies (IgG, IgM, IgE and IgA) were detected by enzyme-linked immunosorbent assay (ELISA) in the sera of hydatid (Echinococcus granulosus) patients from Turkana (Kenya) and the UK. Specific IgG and IgM antibodies predominated in current UK hydatid infections, while all classes of specific antibodies were lower in the Turkana patients. Circulating antigen, detected in 3% polyethylene glycol (PEG) precipitated complexes, using peroxidase conjugated hyperimmune human hydatid IgG (Fab) was more specific in ELISA than either antibody or immune complex assays where peroxidase conjugated anti-human IgG was used. Anti-human immunoglobulin ('rheumatoid' factor) was not detected in hydatid sera. Serum antigen, specific IgM immune complexes and specific IgM antibodies were associated with UK cases of current hydatid infection in contrast to patients with previous histories of hydatidosis. In 3 hydatid patients (from UK) levels of circulating antigen and specific IgM immune complexes rapidly declined within 1-4 months after surgical cyst removal. The detection of specific IgG and antigen in PEG precipitated immune complexes from false-negative/low responder Turkana hydatid sera, suggests that antibody 'mopping' by specific antigen may be occurring. After SDS-PAGE/immunoblotting analysis, antigen of mol. wt 67 000, present in hydatid cyst fluid and protoscoleces, was identified as putative circulating antigen in 3% PEG precipitates of sera from albendazole treated hydatid patients.