Serological differences between acute and chronic schistosomiasis mansoni detected by enzyme-linked immunosorbent assay (ELISA).

Sera from patients with acute and chronic schistosomiasis mansoni, and from laboratory-infected monkeys, were examined by an enzyme-linked immunosorbent assay technique using antigens prepared from eggs, cercariae, and adult worms. Sera from patients with acute schistosomiasis and from monkeys 2 months post-infection reacted more positively to cercarial antigen than to adult worm antigen whereas sera from both patients with chronic schistosomiasis and monkeys infected for longer than 4 months reacted more positively to adult worm antigen. These differential responses to antigen serologically differentiated between acute and chronic schistosome infections.     

血吸虫循环抗原检测的现场试验Ⅱ

制备血吸虫病单克隆抗体诊断试剂盒,将其用于Dot—ELISA,分别检测江苏、安徽流行区急性血吸虫病患者及四川、江西、安徽流行区慢性血吸虫病患者血清中循环抗原。结果不同流行区Dot—ELISA反应的平均“+”值、阳性率和抗原滴度的几何均数不同,循环抗原水平可以反映流行区的疫情。另将Dot—ELISA、ELISA检测四川慢性血吸虫病人治前及治后半年、1年的血清循环抗原及抗体,结果循环抗原在病人治疗后很快下降,治后1年77.8％(35/45)病人的循环抗原转为阴性。循环抗体在病人治疗后也逐渐下降,但速度甚为缓慢。治疗后1年,仅21.7％(10/46)病人转为阴性。上述结果表明,循环抗原能更直接、更真实地反映防治效果。     

Enzyme immunoassay of the level of parasite-specific IgE antibodies in schistosomiasis using the monoclonal antibody BL-IgE 9

Sera from patients with chronic schistosomiasis (Schistosoma mansoni or Schistosoma haematobium) were examined for the presence of parasite specific IgE antibodies by means of ELISA technique using tegument antigen prepared from adult worms of Schistosoma mansoni and using the monoclonal antibody BL-IgE 9. Individuals from tropical countries who had no schistosomiasis and blood donors from GDR were studied for comparison. Significantly higher levels of specific IgE antibody were given by sera from patients with schistosomiasis than by the controls. These differential responses serologically differentiated between patients with chronic schistosome infections and noninfected individuals.     

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。     

多克隆与单克隆抗体夹心ELISA法检测日本血吸虫病患者血清循环抗原

[目的 ]检测血吸虫病患者的循环抗原。 [方法 ]用多抗与单抗夹心 EL ISA法检测日本血吸虫病患者血清循环抗原。 [结果 ]用于检测 15 0例血吸虫病患者的循环抗原 ,其敏感性平均为 84.7% (72 .0 %～ 91.0 % ) ,40例正常人未出现假阳性反应 ,74例其他寄生虫感染者中 ,有 2例并殖吸虫病患者阳性 ,其余均未出现交叉反应。 [结论 ]多抗与单抗夹心 EL ISA法是一种较为理想的循环抗原检测方法。     

日本血吸虫重组信号蛋白14-3-3(rSj14-3-3)及其单克隆抗体用于诊断的价值

目的探讨纯化的日本血吸虫重组信号蛋白14-3-3(rSj14-3-3)及其相应单克隆抗体对日本血吸虫病的诊断价值。方法利用纯化的rSj14-3-3抗原和可溶性虫卵抗原(SEA)间接ELISA方法检测急、慢性日本血吸虫病患者血清中特异性抗体;以纯化的抗rSj14-3-3单克隆抗体包板,以兔抗rSj14-3-3多抗和酶标羊抗兔多抗为检测系统的酶标夹心法检测患者血清中的循环抗原。结果用rSj14-3-3检测急、慢性血吸虫病患者血清中特异性抗体的阳性率分别为100%和933%,用抗rSj14-3-3单克隆抗体检测循环抗原的阳性率分别为100%和956%,两者联合检测的总阳性率分别为100%和978%;经治疗后2、4、6、12个月,抗体转阴率分别为387%、548%、75%、90%,血清循环抗原转阴率分别为177%、419%、55%、85%,两者联合检测的总转阴率分别为452%、63%、85%、90%;对正常人血清的检测未见假阳性反应,与华支睾吸虫病和钩虫病患者血清出现轻微的交叉反应;用SEA检测抗体的阳性率分别为978%和956%,与正常人及华支睾吸虫病和钩虫病患者血清均有不同程度的交叉反应,治疗后2、4、6、12个月阴转率分别为32%、65%、125%、15%。结论rSj14-3-3检测急慢性血吸虫病患者血清中的特异性抗体和利用抗rSj14-3-3单克隆抗体检测循环抗原具有高度的特异性和敏感性,抗原和单抗可规模化     

金标免疫渗滤法(DIGFA)快速检测血吸虫循环抗原的研究

目的　建立一种简便、快速的血吸虫病诊断方法。方法　在已建立的检测血吸虫抗体的ＤＩＧＦＡ基础上,以抗血吸虫重组蛋白ＳＶＬＢＰ多抗为捕捉抗体,金标抗ＳＥＡ多抗为覆盖抗体,建立快速检测病人血清中血吸虫循环抗原的双夹心ＤＩＧＦＡ。结果　用该法检测了急性血吸病人血清３０ 人份和慢性血吸病人血清３８ 人份,其阳性检出率分别为１００ ％ 和５２－６ ％ ;１２０ 人份流行区健康人血清全为阴性;１００ 人份非流行区健康人群血清的阴性符合率为１００％ ,与１０ 例华支睾病人血清无交叉反应;１５ 例肺吸虫病人血清有１ 例阳性,交叉反应率为６－７ ％ ;与双夹心ＥＬＩＳＡ的符合率为９７－４％ ,经χ２ 检验表明两法检测效果无显著性差异。结论　用ＤＩＧＦＡ检测血吸虫循环抗原具有较高的敏感性和特异性,并且方法简便、快速、不需特殊仪器设备,有广阔的应用前景。     

Diagnosis of human schistosomiasis by detection of circulating cathodic antigen with a monoclonal antibody.

Monoclonal antibody (MAb) 5H11 reacted with repeating epitopes on Schistosoma mansoni circulating cathodic antigen (CCA) and detected CCA in sera of Egyptian S. mansoni-infected patients. MAb 5H11 was both capture and biotinylated detection antibody in a sandwich ELISA of trichloroacetic acid-pretreated serum samples. Sera of patients with 7-500 eggs/g of stool were positive by MAb 5H11-CCA sandwich ELISA. Stool egg counts and CCA serum levels correlated (r = .52), and CCA levels decreased by 4 weeks after praziquantel treatment in patients with pretreatment egg counts of greater than or equal to 50/g of stool (P less than .05). Sera of Schistosoma haematobium-infected patients, uninfected individuals, and most patients with other helminths were negative in this assay. The MAb 5H11-CCA sandwich ELISA appears sensitive and specific for immunodetection of active schistosomiasis mansoni and useful for monitoring its chemotherapy.     

斑点-ELISA和独特型/抗独特型抑制试验检测日本血吸虫病患者循环表膜抗原

将血吸虫成虫表膜机原的单克隆抗体8SE4联接辣根过氧化物酶制成结合物,用于直接法斑点-ELISA检测循环抗原。结果48例血吸虫病患者的阳性率为81.3％(39例),24例正常人未出现假阳性反应。检测华支睾吸虫及并殖吸虫病患者各18例,均无交叉反应。本结果证实日本血吸虫病患者血清中有循环表膜抗原存在。直接法斑点-ELISA方法简便,具有较高的敏感性和特异性,可作为诊断日本血吸虫病的补充方法。本文还报告了独特型/抗独特型(8SE_4/抗8SE_4)抑制试验检测循环表膜抗原的初步结果。     

斑点—ELISA和独特型／抗独特型抑制试验...

A monoclonal antibody (designated 8SE) against membrane antigen of adult Schistosoma japonicum labeled with HRP was used in dot-ELISA (direct method) to detect schistosomal circulating membrane antigen. Sera from 48 parasitologically confirmed schistosomiasis cases were tested, 39 (81.3%) were positive. No false positive reaction was found in sera from 24 healthy controls. No cross reaction was detected in sera from clonorchiasis or paragonimiasis in 18 cases each. This results suggest that circulating membrane antigen does exist in patients with schistosomiasis and it might be used as a complementary method for diagnosis of schistosomiasis. A preliminary result of idiotype/antiidiotype interaction inhibition reaction for detecting circulating membrane antigen was also presented.     

Comparative sensitivity and specificity of ELISA and IHA for serodiagnosis of strongyloidiasis with larval antigens

Sera from 68 patients with parasitologically proven strongyloidiasis were tested by the ELISA and IHA tests using larval antigens prepared from Strongyloides stercoralis and Strongyloides ratti. The ELISA using the S. stercoralis antigen detected the greatest number of sero-reactors (83.8%), whereas the IHA using the S. ratti antigen detected the fewest (55.9%). In addition, the S. stercoralis antigen had higher geometric mean titers than the S. ratti antigen in both the ELISA and the IHA tests. Sera from 37 presumed normal individuals also were tested by IHA and ELISA and nonspecific reactions were seen only with the IHA test. When sera from patients with parasitic infections other than strongyloidiasis were tested, the only consistent cross-reactions were with those sera from patients who had occult filariasis and acute schistosomiasis.     

应用特异单克隆抗体测定猪囊尾蚴病人血清和脑脊液中的循环抗原

我们应用特异的抗猪囊尾蚴抗原的单克隆抗体(CCy1),建立了单克隆抗体抑制性ELISA方法以测定囊尾蚴病患者的循环抗原(CAg)。测定灵敏度为ng水平。83例囊尾蚴病患者血清的CAg阳性率为71.1％。其范围为0.16-128μg/ml。对41例囊尾蚴病患者,同时测定了血清和脑脊液中的CAg。单项或两项阳性的总阳性率为90.2％。114例正常人血清、107例其它寄生虫病患者(包括30例包虫病患者)的血清及10例非寄生虫病患者的脑脊液的CAg量均为零。有23例囊尾蚴病患者治疗半年到1年后,有21例抗原浓度为零。另2例分别为0.64μg/ml和1.6μg/ml。表明测定CAg不仅可以确定活动性囊尾蚴感染,而且可考核疗效。     

应用重组信号蛋白14-3-3间接ELISA诊断日本血吸虫病

目的探讨重组日本血吸虫信号蛋白14-3-3(rSj14-3-3)间接ELISA用于血吸虫病免疫诊断的价值。方法表达并利用纯化的rSj14-3-3与成虫抗原(SjAWA)间接ELISA法分别检测51份急性和49份慢性日本血吸虫病患者和50份正常人血清,以上血清标本同时用SEA致敏的间接血凝试验(SEA-IHA)检测。再以3种方法检测30份华支睾吸虫感染者、24份卫氏并殖吸虫感染者和31份钩虫感染者血清,分析其交叉反应性。结果51份急性和49份慢性血吸虫病患者血清的rSj14-3-3抗体阳性率分别为98.0%和91.8%;SjAWA的检出率分别为96.1%和90.0%;IHA的阳性率分别为98.0%和93.9%。经统计学分析,以上结果无显著性差异(P>0.05)。Sj14-3-3间接ELISA、SjAWA间接ELISA和SEA-IHA对于30份华支睾吸虫感染者的交叉反应性分别为13.3%、20.0%和16.7%,24份卫氏并殖吸虫感染者分别为8.3%、12.5%和12.5%,31份钩虫感染者分别为12.9%、16.1%和12.9%。经统计学分析结果也无显著性差异(P>0.05)。结论rSj14-3-3抗原间接ELISA法诊断日本血吸虫病,具有高度的敏感性和特异性,可用于血吸虫病的免疫诊断。     

血吸虫病患者血清中循环25kD表膜抗原的检测

采用细胞杂交瘤技术获得一株血吸虫成虫抗原的单克隆抗体SM 38-3。其靶抗原为成虫表膜,免疫球蛋白型为1gG 2a,能识别血吸虫分子量为25k D的表膜蛋白抗原表位。该单克隆抗体纯化后标记过氧化物酶,用于Dot-ELISA测定慢性血吸虫病人血清115份,阳性反应103份,阳性率为89.6％;测定急性病人血清89份,阳性反应85份,阳性率为95.5％;测定141份正常人血清,阳性反应2份,假阳性率为1.4％。与血吸虫肠相关抗原的检测比较,结果阳性率与抗原滴度均无明显差别。     

Serological studies on schistosomiasis mansoni in the northeast Brazil (I)

Sera from the patients (N = 10) with schistosomiasis mansoni of the hospital of Federal University of Pernambuco, the Schistosoma mansoni egg-positive (N = 51) and -negative (N = 452) inhabitants in Cabo City area, out-patients (N = 37) of the IMIP hospital and Japanese immigrants (N = 127) in Petrolina City area of northeast Brazil as well as Japanese healthy subjects (N = 30) were examined by serological tests including an enzyme-linked immunosorbent assay with antigens prepared from eggs (ELISA-egg) and adult worms (ELISA-adult). The ELISA with egg or adult antigen correctly identified 100% of the uninfected individuals lived in non-endemic area of schistosomiasis. Moreover, when examined cross-reactivity of our ELISA with sera isolated from 78 subjects infected with various intestinal parasitic infections, only one of these sera reacted with the egg and adult antigens. On the examination of 51 sera from the egg-positive subjects, the ELISA-egg revealed the highest sensitivity (98.0%), whereas a large number of false negative reactions of ELISA-adult, Ouchterlony method using adult antigen, circumoval precipitation and immediate intradermal skin test were observed. A low sensitivity of these serologic tests except for ELISA-egg appears to be primarily due to their inability to detect antibody in the sera from egg-positive infantiles. There was no positive correlation between the absorbance values of these two types of ELISA among the sera isolated from ELISA-positive subjects. Rather, by the reactivity of these sera to egg or adult antigen, they could be divided into two subgroups; one reacted more positively with egg antigen and the other with adult antigen. Moreover, it was confirmed that the sera from young subjects (under 20 years old) appear to be highly reactive to the egg antigen than did aged ones. These data suggest that the ELISA with egg antigen, but not with the adult antigen, appears to be useful for the serological survey of schistosomiasis mansoni in the endemic area of northeast Brazil.     

单克隆抗体Dot-ELISA检测血吸虫病循环抗原的研究.Ⅰ.

将血吸虫肠相关阴极抗原的单克隆抗体3D8A联结过氧化物酶后用于直接法Dot-ELISA检测急性、慢性与晚期血吸虫病人血清中循环抗原,阳性率分别为90.6、83.2及30.7％。正常人血清及非寄生虫感染病人血清均未见阳性反应。EPG>100组血清阳性率及抗原水平高于EPG<100。慢性血吸虫病人经吡喹酮治疗后一年,粪检阴性者大部分血清抗原转为阴性;粪检阳性者血清检查仍为阳性。结果表明,肠相关阴极抗原单克隆抗体3D8A能用于Dot-ELISA检测各期病人血清中循环抗原,并能反映病人感染度与药物治疗效果。     

Presence of the schistosome circulating anodic antigen (CAA) in urine of patients with Schistosoma mansoni or S. haematobium infections

We investigated the presence of the circulating anodic antigen (CAA) in the urine of schistosomiasis patients. This genus specific antigen was hitherto demonstrated only in the serum of schistosomiasis patients. The urine of 80 patients with Schistosoma mansoni infections, 33 patients with S. haematobium infections, and 2 patients with mixed S. haematobium and S. mansoni infections were screened by a quantitative enzyme-linked immunosorbent assay (ELISA). CAA was demonstrated in 81% of those with intestinal schistosomiasis and in 97% of those with urinary schistosomiasis. CAA titers were less than 1:0.2-1:51.2. Results were compared with circulating cathodic antigen (CCA) titers in urine obtained in an indirect hemagglutination assay (IHA). CCA was generally not detectable in the urine of patients with S. haematobium infection, but was demonstrated in the urine of 85% of the patients with S. mansoni infection. Both CAA titers and CCA titers correlated positively with the number of S. mansoni eggs excreted in the feces, but CAA titers did not show a significant correlation with the number of S. haematobium eggs in urine. Both antigen titers showed a moderate correlation with the serum CAA level in schistosomiasis mansoni. The discovery of CAA in the urine of the majority of schistosomiasis patients tested suggests the use of urine samples for non-invasive immunodiagnosis of the disease.     

Presentation of autoantibody to proliferating cell nuclear antigen in patients with chronic hepatitis B and C virus infection

OBJECTIVES: To study the association of antibodies to proliferating cell nuclear antigen (PCNA) in patients with chronic hepatitis B (HBV) and C (HCV) virus infection. METHODS: Sera from 243 patients with chronic HBV infection; 379 patients with chronic HCV infection; 80 patients with systemic lupus erythematosus (SLE); 28 patients with rheumatoid arthritis; 15 patients with Sjogren's syndrome; eight with polymyositis; eight with primary biliary cirrhosis; and 33 healthy control subjects were tested for the presentation of anti-PCNA antibodies by enzyme linked immunosorbent assay (ELISA) and immunoblotting using recombinant PCNA as antigen. The distribution of immunoglobulin isotypes of anti-PCNA antibody was measured by ELISA assay. RESULTS: By ELISA, anti-PCNA antibodies were detected in 30 (12.3%) patients with chronic HBV infection, 71 (18.7%) patients with chronic HCV infection, and five (6.3%) patients with SLE. The inhibition of binding with these sera by purified PCNA was shown to exceed 71%. By immunoblotting, the frequency of anti-PCNA in patients with chronic HBV and HCV infection was 17 of 243 (7%) and 41 of 379 (11%), respectively. Absorption studies on indirect immunofluorescence showed the typical nuclear speckled staining pattern by anti-PCNA sera was abolished by preincubation of sera with PCNA. Anti-PCNA antibody was not detected in sera from patients with autoimmune diseases except SLE. Anti-PCNA antibodies in patients with chronic HBV and HCV infection were predominantly IgG. CONCLUSION: These data suggest that anti-PCNA antibody are also present in patients with chronic HBV and HCV infection. Anti-PCNA antibody may not be specific for SLE.     

金标渗滤法快速检测血吸虫循环抗原的现场应用研究

将抗重组蛋白ＳＶＬＢＰ－ＩｇＧ用于ＤＩＧＦＡ检测２２２１例血清中血吸虫循环抗原,其中急性血吸虫病人血清７０份,慢性血吸虫病人血清３０７份,循环抗原检出率分别为１００％和６８．４％。非流行区健康人血清２００份,未见阳性反应。除肺吸虫外,与其它寄生虫或非寄生虫感染病人血清无明显交叉反应。用ＤＩＧＦＡ单盲法检测２批血清,结果显示对急性血吸虫病人的敏感性均为１００％;对慢性血吸虫病人的敏感性分别为６９．４     

Long-term persistence of hepatitis C virus antibodies in a single source outbreak.

The occurrence of antibodies to hepatitis C virus (HCV) was investigated in 81 patients who developed hepatitis non-A, non-B (HNANB) after parenteral administration of contaminated immunoglobulin to prevent Rh sensitization. Sera from 74 of the 81 patients (89.9%) were anti-HCV positive at either 6-12 months or 9-10 years after administration of immunoglobulin. Sera were not available from any patients at either of the times: however, 52 of 56 sera (92.9%) were anti-HCV positive 6-12 months after use of immunoglobulin, and anti-HCV was present in 45 of 65 sera (69.2%) 9-10 years after immunoglobulin treatment. Of the latter, only two of 13 (15.4%) sera from patients who recovered from hepatitis were anti-HCV positive, whereas 43 of 52 patients (82.7%) with chronic disease were anti-HCV positive. The ELISA using a recombinant antigen was found a good detector as marker for a HCV infection because 90% of patients infected by a common source became anti-HCV positive. However, 10 years after infection most patients who did not develop chronic disease no longer had detectable antibodies.