日本血吸虫不同发育阶段抗原检测IgG及IgG_4的疗效考核价值

目的 :建立具有疗效考核价值的抗原 -抗体检测系统。方法 :用日本血吸虫不同发育阶段抗原 (AWA、AMA、AESA、SEA、SIEA)检测治疗前及治疗后不同时间慢性血吸虫病人血清中特异性IgG及IgG4 并测定肺吸虫病人、肝吸虫病人和健康人血清的交叉反应性。结果 :不同发育阶段抗原 -IgG检测系统间对慢性血吸虫病人、肺吸虫病人、肝吸虫病人和健康人血清测试的阳性率无显著性差异 (P >0 .0 5 ) ,慢性血吸虫病人治疗后 12个月IgG的阴转率以SIEA最高 (90 .0 % ) ,其次为AMA(77.5 % ) ,最低为AESA(2 0 .0 % ) ;检测慢性血吸虫病人治疗前的特异性IgG4 的阳性率AWA最高 (85 .0 % ) ,其次为AMA(80 .0 % ) ,最低的为SIEA(37.5 % ) ,治疗后 12个月的IgG4 阴转率SIEA最高 (97.5 % ) ,其次为AMA(95 .0 % )和AWA(85 .0 % ) ,最低的为SEA(6 2 .5 % )。结论 :AMA -IgG、SIEA -IgG、AWA -IgG4 和AMA -IgG4 检测系统具有较好的疗效考核价值 ,且以SIEA -IgG和AMA -IgG4 两个检测系统最好。     

Detection of a circulating SJ 70 antigen in human schistosomiasis japonica using a monoclonal antibody

A double antibody enzyme-linked immunosorbent assay (sandwich ELISA) was used for detection of a circulating antigen from human Schistosoma japonicum infections. This assay involves the use of poly clonal rabbit anti-schistosoma japonicum soluble egg antigen (SEA) antiserum to bind circulating antigen and a monoclonal antibody (MCAb-H4) to identify and quantify this antigen. Sera from 108 S. japonicum infected patients (acute and chronic) were tested. Sera from 93 of 95 (98%) patients with chronic infection were positive for this antigen; sera from 12 of 13 patients (93%) with acute infections were also positive. Antigen was not detectable in control human sera. Sera from 35 chronic schistosomiasis patients were collected between 6 and 12 months after paraziquantel treatment. Circulating antigen was not detectable in the sera from 33 of these patients (94%) and was dramatically reduced in the other two patients.     

曼氏血吸虫与日本血吸虫交叉免疫原性的血清学实验研究

以曼氏血吸虫的虫卵和成虫免疫家兔后所产生的特异性抗体,可用以日本血吸虫虫卵、尾蚴和成虫为抗原分别进行的COP,CHR和ELACIEP测出,表明两种人体血吸虫存有显著的交叉抗原成分。应用此种血清交叉反应性,以检测抗异种人体血吸虫的抗体,似有效而可取的,可用以辅助诊断援外回国人员是否感染国外人体血吸虫病。     

日本和曼氏血吸虫抗原在间接血凝试验中与不同血吸虫病人血清的交叉反应

用间接血凝试验(IHA)比较不同抗原SjSEA、SmAWA和SmBW与日本、曼氏和埃及血吸虫病人血清的交叉反应。结果显示:3种抗原致敏的绵羊红细胞(SRBC)与其相应的血吸虫病人血清反应率分别为100％(SjSEA),96％(SmAWA)和98％(SmBW);与不同种血吸虫病人血清的交叉反应率在79％～95％,其中以SjSEA最高。结果提示:SjSEA除可用于诊断日本血吸虫病外,也可作为诊断曼氏和埃及血吸虫病的候选抗原。此外,本试验还发现IHA抗体滴度与病人排出的虫卵数量无相关关系。     

Characterization of cDNA from the miracidial antigen family of Schistosoma japonicum (Chinese strain)

Objective To identify the egg antigens related to the formation of hepatic granulomas and fibrosis of Schistosomiasis japonica.Methods The egg cDNA library of Schistosoma japonicum (S. japonicum) was constructed and screened by immunological methods with the pooled sera of advanced schistosomiasis patients. The inserted foreign DNA fragments of positive clones were sequenced. The sequence data were analyzed using Wdnasis 2.5 and compared with Genebank data using blast software.Results Eighty-one clones containing recombinant DNA fragments were obtained from the egg cDNA library of S. japonicum by immunological screening. The DNA sequences of all clones belonged to the miracidial antigen family. The longest cDNA fragment was 1604 bp, which contained an open reading frame of 351 bp, which encoded a protein of 1 2913.35 daltons.Conclusion The cDNA sequence of the miracidial antigen of S. japonicum (Chinese strain) was obtained for the first time.     

rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值研究

目的研究rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值。方法用rSj26-Sj32融合蛋白和日本血吸虫成虫抗原(SjAWA)Immunogold-Dipstick法检测慢性日本血吸虫病患者血清,同时以华支睾吸虫病、卫氏并殖吸虫病、泡型棘球蚴病、囊性棘球蚴病、乙型肝炎、肺结核患者和健康人血清作为对照,比较两种抗原检测抗体效果的差异。结果该法检测慢性日本血吸虫病的敏感性和特异性分别为92.50%和97.67%,诊断该病的阳性预告值、阴性预告值及诊断效率分别为97.37%、93.33%和95.18%,并且与其他疾病患者血清均无交叉反应。结论 rSj26-Sj32-Immunogold-Dipstick试剂可用于慢性日本血吸虫病的免疫诊断。     

日本血吸虫成虫脲溶42 kD蛋白的分离及其初步诊断应用

目的 :分离日本血吸虫成虫脲溶抗原中有免疫学活性的抗原分子 ,探讨其诊断血吸虫病的效果。方法 :利用SDS PAGE及免疫印迹法分析日本血吸虫成虫脲溶抗原 ,采用电泳层析法分离有免疫活性的 4 2kD蛋白 ,ELISA分析 4 2kD蛋白诊断血吸虫病的灵敏度和特异性。结果 :电泳层析获得了具有免疫学活性的 4 2kD蛋白 ;分离的 4 2kD蛋白诊断血吸虫病的灵敏度和特异性分别为 88.0 9%和 97.0 6 %。结论 :日本血吸虫成虫脲溶 4 2kD蛋白是一种日本血吸虫的血清学抗原 ,可用于血吸虫病诊断     

4种抗原用于斑点免疫金银染色试验诊断日本血吸虫病的比较

采用斑点免疫金银染色(Dot-IGSS)试验对4种日本血吸虫抗原的敏感性及特异性进行比较。结果显示粗提成虫抗原(AWA)、尿素溶解性成虫抗原(JWU)、硫酸铵沉淀成虫抗原(AW)和可溶性虫卵抗原(SEA)检测40例日本血吸虫病人血清抗体的敏感性均达100.0％,40例正常人血清的可疑阳性反应分别为2.5％、0、2.5％和0,40例肝吸虫病人血清的可疑交叉率分别为2.5％、0、2.5％和0,表明在高敏感的IGSS检测系统中,粗提成虫抗原的敏感性和特异性与部分纯化者和虫卵抗原无显著性差异(P>0.05)。     

Establishment and characterization of a monoclonal antibody against egg antigens of Schistosoma japonicum

A monoclonal antibody (N29B3) against egg antigens of Schistosoma japonicum was obtained by hybridoma technique N29B3 was of IgG3 isotype. The molecular weight of its target antigen was 141 KD. IIFA with section of liver from infected rabbit showed that N29B3 was located on surface of miracidium and egg shell. IIFA and ELISA also showed that N29B3 was not reactive with antigens of differential developmental stages of S. japonicum and antigens of other trematodes. The data indicated that N29B3 has specificity for egg antigens. In western blotting, sera from chronic infected human and mice and from acute infected mice (30 and 45 days) all recognized 141 KD molecule. These results suggest that 141 KD molecule has potential capacity for early diagnosis of schistosomiasis.     

日本血吸虫循环免疫复合物中抗原的鉴定

用低浓度ＰＥＧ（ＭＷ６０００）提取的日本血吸虫感染兔血清中循环免疫复合物（ＣＩＣ）直接免疫家兔，制备ＣＩＣ兔血清。采用酶联免疫印迹技术（ＥＩＴＢ）分析鉴定日本血吸虫ＣＩＣ中抗原成份。结果表明，ＣＩＣ中成虫及其排泄分泌物源性抗原成份有１７种，虫卵源性抗原成份有５种。且ＩＦＡＴ方法分析ＣＩＣ中抗原成份定位于成虫表膜及肠腔上皮。     

日本血吸虫成虫67kDa分子抗原的纯化及其对血吸虫病的诊断和疗效考核

目的 为检测日本血吸虫成虫67kDa分子抗原(SjAWA67)对血吸虫病的诊断及疗效考核价值。方法 通过SDS-PAGE和电渗方法,从日本血吸虫成虫抗原中分离纯化出67kDa分子抗原,并用该抗原包被酶标反应板微孔,进行ELISA检测。结果 SjAWA67的纯度已达到电泳纯和免疫纯,对急、慢性血吸虫病患血清的捡出率分别为100％和95％,与正常人血清、肝吸虫病和肺吸虫病患血清均未出现明显的交叉反应,44例血吸虫病患治疗后3、6和12个月后的阴转率分别达45．5％、75．0％和90,9％。结论 SjAWA67分子抗原具有较好的疗效考核价值和现场应用前景。     

日本血吸虫重组信号蛋白14-3-3对日本血吸虫病的诊断价值

目的 探讨纯化的日本血吸虫(中国大陆株)重组信号蛋白14—3—3(rSjl4—3—3)诊断血吸虫病的价值。方法 利用纯化的rSjl4—3—3抗原与成虫抗原，间接EUSA法检测急、慢性日本血吸虫病患者血清。结果 急、慢性血吸虫患者血清rSjl4—3—3抗体检出率为100％和92．0％，与正常人血清未出现交叉反应；抗AWA抗体检出率为98．0％和94．0％，与正常人有7．3％的交叉反应。结论 rSjl4—3—3为抗原诊断日本血吸虫病显示出高度的敏感性和特异性，具有实用价值。     

用日本血吸虫成虫抗原ELISA检测血吸虫病抗体的现场应用

用日本血吸虫成虫抗原(AAg)和虫卵抗原(EAg)ELISA检测血吸虫病疫区粪检阳性病人134例,其阳性率分别为99.25％和98.51％。134例中45例儿童和89例成人用AAg检测,其阳性率分别为100％和98.88％,而用EAg检测的阳性率分别为97.78％和98.88％。AAg和EAg检测26例正常人均为阴性。结果表明:AAg与EAg的检出率以及儿童与成人的检出率均相似。     

PRELIMINARY STUDIES ON SHARED ANTIGENS AMONG CERCARIA,ADULT WORM AND EGG OF SCHISTOSOMA JAPONICUM

The pooled sera of inbred C57BL/6 mice in- fected with Schistosoma japonicum, the antisera against gut-associated antigen (GAA) from artificially immunized rabbits (RAGAA20) and the antisera against soluble egg antigen (RASEA) purified by im- munoaffinity chromatography were tested with cer caria-stuck-slides, adult worm sections and liver-egg sections by immunohistochemical studies with peroxi- dase-linked protein A (PPA method). In mice anti body against eggs was detected before oviposition. Eggs containing late developmental stage of embryo started to secrete SEA 37 days after infection when the titer of the antibody against eggs reached l:256. GAA20 and SEA were proved to be at the same location within mature eggs, and RAGAA20 0nly reacted with eggs exuding SEA. RASEA reacted strongly with the membrane of cercaria. The results showed that S japonicum SEA shared its antigens with GAA and membrane-asso. ciated antigens (MAA). GAA might play a role of prcsensitizing in egg granuloma formation.     

日本血吸虫不同发育阶段与旋毛虫抗原交叉性的研究

运用ＥＩＴＢ技术分析日本血吸虫（Ｓｊ）尾蚴、肝期童虫及雌雄成虫与旋毛虫（Ｔｓ）肌蚴抗原的交叉性。结果显示：Ｓｊ尾蚴、肝期童虫、成虫各阶段不同程度地被抗－ＴｓＳＡＲＳ，ＴｓＩＲＳ识别，其识别的抗原分子量在１５￣１００（尤５０￣７０）ｋＤ之间，两种抗血清识别的带型基本相同。所识别的各期Ｓｊ抗原中以尾蚴抗原最多，次为肝期童虫。３０天成虫仅雌虫抗原与上述两种抗血清在９７ｋＤ处显出一条弱带。抗－Ｓｊ♂ＡＲＳ     

旋毛虫肌蚴可溶性抗原与日本血吸虫病患者血清的交叉反应

应用ＥＬＩＢ技术观察旋毛虫肌蚴抗原（ＴｓＭＬＳＡ）与６种寄生虫病人血清的交叉反应，结果表明：ＴｓＭＬＳＡ中３１～１００ＫＤａ，内的蛋白大部分被急性血吸虫颊人血清所识别，而慢性血吸虫病人血清仅识别其中的４４／４５，５１／５３，６２／６４及１００ＫＤａ蛋白，小于６０ＫＤａ者可与丝虫病，钩虫病，蛔虫病和肝吸虫病人血清呈不同程度的交叉反应，以前三者为最明显，４５ＫＤａ蛋白可被部分正常人血清识别，提示，旋毛     

血吸虫病间接红细胞凝集试验的标准化研究——抗原的筛选及标准化

分别用日本血吸虫的虫卵可溶性抗原(SEA)、成虫尿素提取抗原(AUA)、成虫表皮膜抗原(ATA)以及SEA与AUA、SEA与ATA的混合抗原致敏绵羊红细胞,并用于检测198例日本血吸虫病人血清,IHA的阳性符合率分别为95.45％、84.67％、90.23％、98.08％、96.25％;检测100例健康人血清,结果全部阴性;检测191例肺吸虫病,旋毛虫病、华枝睾吸虫病和囊虫病患者血清,其总交叉反应率分别为13.61％(SEA)、3.14％(AUA)、4.71％(ATA)、8.37％(SEA+ATA)和3.66％(SEA+AUA)。结果表明:SEA+AUA的制备方法简单,并具有较高的敏感性和特异性。     

三联抗原酶免疫试验诊断日本血吸虫病

用血吸虫成虫—卵—尾蚴三联抗原EIA,检测类卵阳性患者血清94份、非疫区有疫水接触史皮试阳性者血清190份、基本消灭血吸虫病地区人群血清205份,阳性率分别为100％、27.9％及10.2％。69份肝吸虫病人血清,虫卵抗原EIA假阳性率为1.4％;与72份肺吸虫病人血清未显交叉反应;153份献血员血清,其虫卵及尾蚴抗原EIA假阳性率分别为1.3％和2.6％。用三联抗原EIA、成虫粗抗原ELISA、COP及CHR检测94份粪卵阳性患者血清,仅卵抗原EIA 阳性率明显高于COP;成虫及尾蚴抗原 EIA的阳性率分别和成虫粗抗原 ELISA、CHR 的阳性率相同。     

日本血吸虫循环免疫复合物中抗原的鉴定

用低浓度ＰＥＧ（ＭＷ６０００）提取的日本血吸虫感染兔血清中循环免疫复合物（ＣＩＣ）直接免疫家兔，制备抗ＣＩＣ兔血清。采用酶联免疫印迹技术（ＥＩＴＢ）分析鉴定日本血吸虫ＣＩＣ中抗原成份。结果表明，ＣＩＣ中成虫及其排泄分泌物源性抗原成份有１７种，虫卵源性抗原成份有５种。用ＩＦＡＴ方法分析ＣＩＣ中抗原成份定位于成虫表膜及肠腔上皮。     

斑点酶联免疫吸附试验检测血吸虫病

用成虫、卵及尾蚴抗原做斑点酶联免疫吸附试验(Dot—ELISA)检测96份血吸虫患者血清,阳性率分别、为70.8%、91.7%及81.3%。三种抗原 Dot—ELISA 的互补阳性率为93.8%。而对50份献血员及38份肺吸虫患者血清进行检测都是阴性。Dot—ELHSA 的重现性及稳定性好,抗原用量少、简单、出结果快。