动物致伤人群接种狂犬病疫苗后血清学与流行病学效果的初步分析

目的观察就诊的动物致伤者,分别全程足量接种两种剂型狂犬病疫苗后,其血清中抗体阳转率和疫苗保护率。方法2000—2001年就诊者全程接种浓缩苗,2002—2003年的全程接种精制纯化(vero细胞)苗。严重致伤者,于0、3天接种加倍量疫苗,并于0天的同时,对其伤口周围用抗狂犬病血清(40IU/kg)进行浸润注射。联合使用抗血清者,完成全程5次注射后第15、75天各加强1针疫苗。接种结束后15—20天,以间接混合酶标法检测血清中抗狂犬病病毒抗体。期内经疫情报告系统,收集当年观察人群狂犬病病例,计算两接种组人群的发病率,并分别与当年未接种疫苗的动物致伤人群比较,以估算疫苗保护率。结果4年中该人群抗体总阳性率为95.60%。接种精制纯化苗组的阳性率为96.37%,高于浓缩苗组的94.83%。狂犬病疫苗保护率,精制苗1年和2年的均为100.00%;浓缩苗1年的为95.46%,2年、3年和4年的为90.91%。结论动物致伤人群规范处理局部伤口后,及时全程接种狂犬病疫苗,其抗体阳性率较高。国产精制纯化苗接种人群的抗体阳性率和1~2年的疫苗保护率高于浓缩苗。从我国狂犬病防治工作实际出发,提示国产精制纯化苗需尽快降低成本,以巩固和提高其接种率。     

两种试剂盒快速检测斯氏狸殖吸虫抗体的研究

目的观察肺吸虫病金标渗滤试剂盒(DIGFA kit)和酶联免疫吸附试验(ELISA)试剂盒检测斯氏狸殖吸虫抗体的敏感性和特异性。方法采用DIGFA kit和ELISA检测斯氏狸殖吸虫病流行区人群和病鼠血清抗体。结果 DIGFA和ELISA试剂盒检测斯氏狸殖吸虫病流行区人群和病鼠血清特异性抗体阳性率均为5.08%(54/1062)和100%(40/40);用两种试剂盒检测正常大鼠、旋毛虫病大鼠、血吸虫病兔和蛔虫病病人血清,除1例血吸虫病兔血清DIGFA出现阳性反应外,其他血清两种方法均为阴性。大鼠感染斯氏狸殖吸虫后第2周用两种方法检测抗体,阳性率分别为20.00%(8/40)和27.50%(11/40),第4周阳性率均达100%,并持续至第8周。结论 DIGFA试剂盒具有与ELISA试剂盒相同的敏感性和特异性,且有操作快速、简便、直观等特点,适合在基层和斯氏狸殖吸虫病流行地区推广使用。     

检测抗Sj23HD IgG在监测预警哨鼠血吸虫感染早期诊断中的价值

目的探讨检测抗日本血吸虫23ku膜蛋白大亲水肽段(Sj23HD)抗体IgG对血吸虫感染监测预警哨鼠进行早期诊断的价值。方法在潜在高风险血吸虫感染水域投放哨鼠,第一批哨鼠分别于回收后第14、35d采血,分离血清,第35d解剖查虫,第二批哨鼠分别于第21、50d采血,分离血清,第50d解剖查虫。以重组Sj23HD为抗原,通过常规ELISA和Western blot方法检测哨鼠血清抗Sj23HD抗体IgG,并与解剖查虫结果进行比较。结果 ELISA检测第一批预警哨鼠回收后第14d特异性抗体IgG阳性率为0.4%,第35d阳性率为3.7%;Western blot检测阳性率分别为1.7%和3.7%;第35d解剖,成虫阳性率为0.92%。ELISA检测第二批预警哨鼠回收后第21d特异性抗体IgG阳性率为2.4%,第50d阳性率为12.5%,Western blot阳性率分别为为5.2%和12%。第50d解剖,成虫阳性率为8.2%。两批哨鼠回收后14d、21d,Western blot法阳性率均比常规ELISA法高,两种免疫学方法的阳性率与解剖查虫法阳性率比较差异均有统计学意义(P<0.05)。结论抗Sj23HD抗体IgG ELISA和Western blot检测的敏感性高于解剖查虫法,能提前哨鼠预警的时间,提高预警效果;Western blot法敏感性和特异性均显著高于ELISA法,更具实用价值。     

国产冻干无佐剂Vero细胞狂犬病疫苗接种效果观察

目的观察狂犬病暴露前人群应用国产冻干无佐剂Vero细胞狂犬病疫苗小剂量皮内注射的接种反应及其免疫原性。方法选择犬密切接触人群,按照知情自愿的原则,对244名Ⅰ级狂犬病暴露者接种国产纯化冻干无佐剂Vero细胞狂犬病疫苗(PVRV),按照随机、单盲的原则分成两组,A组122人,分别于第0、7、28 d皮内注射,每个部位0.1 ml;B组122人,分别于第0、7、28 d全量(0.5 ml)PVRV三角肌注射。观察受种者疫苗注射部位皮内及肌肉局部和全身反应,采用间接ELISA法检测血清抗体。结果皮内注射组局部红肿、硬结、疼痛、瘙痒,发生率分别为1.09%、0.27%、0.27%和10.66%,肌肉注射组分别为0.82%、0.27%、1.64%和0.82%。皮内注射组发热、皮疹、头痛、疲劳乏力等全身反应发生率分别为0.27%、0.27%、0.55%和0.82%,肌肉注射组分别为0.55%、0.27%、0.55%和0.82%。不良反应常发生在第1、第2次注射之后。皮内注射(ID)组、肌肉注射(I M)组接种狂犬病疫苗后42 d抗体阳转率分别是94.21%和95.08%,差异无统计学意义(P0.05)。结论国产冻干Vero细胞狂犬病疫苗免疫原性良好,皮内、肌肉注射不良反应轻微。对暴露前人群小剂量皮内注射国产冻干无佐剂Vero细胞狂犬病疫苗是可行的。     

狂犬病疫苗接种982例效果观察

采用ELISA法，对982例接种狂犬疫苗者血清进行狂犬病毒抗体测定，平均抗体阳性率为90．12％。个体年龄差异对狂犬病毒抗体形成有一定的影响。全程、规范注射狂犬病纯化疫苗是预防狂犬病发生的最有效的方法。     

间接ELISA检测弓形虫感染鼠NTPase-Ⅱ抗体的研究

目的建立检测小鼠血清中刚地弓形虫NTPase-Ⅱ特异性IgGI、gM抗体的间接ELISA法,探讨用于弓形虫感染早期检测的应用价值。方法以刚地弓形虫NTPase-Ⅱ融合蛋白为包被抗原建立间接ELISA法,动态检测刚地弓形虫实验感染鼠血清NTPase-Ⅱ特异性IgGI、gM抗体变化。试验设脑囊虫病、血吸虫病及肺吸虫病病人血清对照。结果小鼠血清中的抗NTPase-Ⅱ抗体IgM于弓形虫感染后第3 d开始出现阳性反应,IgG抗体于感染后第7 d开始出现阳性反应,脑囊虫病、血吸虫病及肺吸虫病病人血清特异性IgG阳性率分别为0、1.92%和0。结论间接ELISA检测弓形虫感染鼠血清NTPase-Ⅱ抗体的敏感性和特异性较高,有望应用于人感染弓形虫的早期检测。     

5119例狂犬病疫苗全程免疫接种者血清抗体水平检测结果分析

目的了解狂犬病疫苗全程免疫后抗体产生情况及其影响因素,为狂犬病疫苗预防接种工作提供依据。方法采用ELISA法对5119例于我院全程接种狂犬病疫苗者进行抗狂犬病病毒抗体检测并分析结果。结果5119份血清标本中抗体阳性5035份,总阳性率98．36％。其中男性及女性阳性率分别为97．91％、98．75％;P〈0．05。0—19岁、20～59岁及〉60岁者抗体阳性率分别为99．3％、98、4％、96．3％,两两比较,P均〈0．05;即随着年龄增大,接种疫苗后抗体阳性率呈下降趋势。结论性别对狂犬疫苗接种后抗体的产生有影响;随着年龄的增长,注射疫苗后抗体阳性率逐渐下降。对接种后抗体阴性者应再次进行全程免疫。     

金标免疫渗滤法诊断试剂盒诊断血吸虫病的价值研究

目的　评价金标免疫渗滤法诊断试剂盒（DIGFA-kit）用于血吸虫病血清流行病学调查的价值。方法　采用DIGFA与ELISA单盲平行检测各期血吸虫病病人血清、非流行区人群血清、疫区人群血清和血吸虫病已控制地区人群血清共2304份。结果　DIGFA检测血吸虫抗体的敏感性为96.8%，对非流行区健康人群的特异性为100%；该法检测疫区居民血清508份，抗体阳性检出率为35.6%，在Kato法粪检阳性的81人中，其中抗体阳性79人，阳性符合率为97.5%；检测血吸虫病已控制地区人群血清，抗体阳性率为4.6%；根据DIGFA的敏感性和对非流行区人群的特异性，计算其Youdens指数为0.926。上述各项结果与ELISA比较，差异无显著性（P>0.05）。结论　DIGFA与ELISA有相似的敏感性和特异性，且方法简便、快速，胶体金标记物制备容易，试剂稳定，运送方便，在现场和临床诊断更具有优越性。     

NP30/抗体检测试剂盒在云南大山区型血吸虫病流行区的现场应用

目的用NP30/抗体检测试剂盒在大山区型血吸虫病流行区筛查病人,评价其应用价值。方法在云南省大山区型血吸虫病流行区,选择3个自然村作为现场,随机抽取10～70岁的村民作为调查对象,进行血吸虫病病史个案调查;收集粪便,作毛蚴孵化检查;静脉采血,分离血清,分别采用NP30/抗体检测试剂盒和酶联免疫吸附试验试剂盒（SEA/ELISA法）检测特异性IgE和总IgE抗体。结果3个村共筛查村民506人,粪便毛蚴孵化阳性164例,阳性率为32.41%;NP30和SEA/ELISA的阳性率分别为58.30%和75.10%。在164例粪检阳性血清样本中,NP30和SEA/ELISA的阳性符合率分别为87.80%和84.76%。在342例粪孵阴性血清样本中,NP30和SEA/ELISA的阳性率分别为44.15%和70.47%。结论NP30/抗体检测试剂盒具有较高的敏感性和特异性,可用于大山区型血吸虫病流行区病人筛查。     

应用捕获ELISA检测肾综合征出血热患者尿液中特异性IgM抗体

目的检测肾综合征出血热患者尿液中特异性IgM抗体,研究早期诊断方法.方法用MacELISA法检测不同病日HFRS病人尿液中特异性IgM抗体.结果HFRS病人尿夜中特异性IgM抗体总阳性率为76.47%,第3病日即可出现阳性,7-9病日阳性率可达83.87%,与同期HFRS病人血清抗体检测对比无显著性差异(P＞0.05).结论用MacELISA检测HFRS尿液中特异性IgM抗体敏感性高,特异性强,适合早期诊断.     

An ELISA suitable for the detection of rabies virus antibodies in serum samples from human vaccinated with either cell-culture vaccine or suckling-mouse-brain vaccine.

An indirect ELISA for determination of post-vaccination rabies antibody was applied. Purified rabies virus was used as antigen to coat plates, and staphylococcal protein A linked with horseradish peroxidase was used for detecting IgG antibody in human sera. Sera from humans, vaccinated with cell-culture vaccine or suckling-mouse-brain vaccine, were examined. ELISA results were compared to those obtained from the virus neutralization test. The mean and standard deviation of OD were determined for 126 negative sera (pre-vaccination) and for 73 sera from vaccinated persons showing antibody titers lower than 0.5 IU/ml. Results were defined as ELISA -positive, -negative or -doubtful. Establishment of a doubtful region reduced the number of sera otherwise classified as positive (false-positive sera). In this way, the sensitivity, specificity and agreement values were respectively 87.5%, 92.4% and 88.5%. No significant differences were observed in these values when the group vaccinated with cell-culture vaccine and the group vaccinated with suckling-mouse-brain vaccine were compared. It was shown that much of the disagreement between the values obtained by neutralization test and ELISA occurred in sera obtained at the beginning of the immunization process, and was probably due to the presence of IgM in the serum samples, detected only by the former test. This ELISA method can be used as a screening test in rabies laboratories regardless of the kind of vaccine used for immunization.     

Antibody response in cattle after vaccination with inactivated and attenuated rabies vaccines

Despite the absence of current official reports showing the number of cattle infected by rabies, it is estimated that nearly 30,000 bovines are lost each year in Brazil. In order to minimize the important economic losses, control of the disease is achieved by eliminating bat colonies and by herd vaccination. In this study, we compare the antibody response in cattle elicited by vaccination with an attenuated ERA vaccine (AEvac) and an inactivated-adjuvanted PV (IPVvac) vaccine. The antibody titers were appraised by cell-culture neutralization test and ELISA, and the percentage of seropositivity was ascertained for a period of 180 days. IPVvac elicited complete seropositivity rates from day 30 to day 150, and even on day 180, 87% of the sera showed virus-neutralizing antibody titers (VNA) higher than 0.5IU/ml. There were no significant differences between the VNA titers and seropositivity rates obtained with IPVvac in the two methods tested. AEvac, however, elicited significantly lower titers than those observed in the group receiving inactivated vaccine. In addition, the profiles of antirabies IgG antibodies, evaluated by ELISA, and VNA, appraised by cell-culture neutralization test, were slightly different, when both vaccines were compared.     

Antibody responses in Japanese volunteers after immunization with yellow fever vaccine

To monitor the development of specific and cross-reactive antibody response in twenty Japanese volunteers after vaccination with live yellow fever vaccine. Serum samples were collected on various days after vaccination and examined for hemagglutination inhibition (HI) antibodies against yellow fever virus (YFV), Japanese encephalitis virus (JEV) and dengue virus (DV), neutralizing antibodies against YFV and JEV, and IgM antibodies against YFV. None of the volunteers had been previously immunized with this vaccine. Fifteen of 20 had pre-vaccinated with JEV 7 to 40 years before. Ten of the 20 had neutralizing antibodies against JEV before immunization. None of the 20 had detectable antibodies against YFV or DV before vaccination. On day 10th after the vaccination, neutralizing antibodies to YFV were detected in 6 of 19 volunteers and IgM antibodies against YFV were detected in 7 of 19. On day 14th, HI, neutralizing, and IgM antibodies against YFV were detected in all the tested sera. Neutralizing antibodies against JEV were developed in 2 volunteers and HI antibodies against JEV were increased in 3 of 6 volunteers respectively. On day 29th, cross-reactive HI antibodies for JEV and DV were detected in all the tested sera. The results indicate that YF vaccine induces YFV-specific antibodies in all the tested volunteers and that it also induces HI antibodies cross-reactive for JEV and DV. The YF vaccine has a strong immunogenicity because it is a live vaccine, and induces antibody against YFV predominantly. The international certificate of yellow fever vaccination becomes valid 10 days after vaccination. On day 14th after vaccination, we detected neutralizing antibodies against YFV from all tested volunteers, however, only 6 of 19 volunteers had detectable neutralizing antibody on the 10th day after vaccination. Therefore, the vaccine may not be perfectly effective on day 10th after the vaccination.     

Antibody responses to human diploid cell vaccine for rabies with and without human rabies immune globulin

One hundred one volunteers with no exposure to rabies were given human diploid cell vaccine (HDCV) for rabies with or without 20 international units of human rabies immune globulin (HRIG)/kg of body weight to evaluate schedules for therapy with HDCV and HRIG after exposure. All of the volunteers who received three or more doses of HDCV alone or four or more doses of HDCV with HRIG developed high titers of neutralizing antibodies by day 35, which persisted for at least 60 days. By day 7, of the 61 volunteers given HRIG and HDCV, 53% had neutralizing antibodies by a mouse neutralization test and 67% had neutralizing antibodies by a rapid fluorescent focus inhibition test. Similar antibody levels were found in volunteers given HRIG alone, a finding which suggests that low or undetectable early titers after administration of HDCV and HRIG were due to inadequate HRIG dosage rather than any interaction between the passive antibody (HRIG) and the vaccine antigen. These results suggest that trials with 30 or 40 international units of HRIG/kg in combination with HDCV are warranted.     

鹦鹉热衣原体重组主要外膜蛋白诱导肉鸡免疫应答观察

用鹦鹉热衣原体 (Chlamydiapsittaci,Cps)重组主要外膜蛋白 (RecombinentMajorOuter Membraneprotein ,r MOMP)制作成油佐剂疫苗 ,经颈部皮下免疫 5d肉鸡。在免疫前 1d、免疫后 14d、2 1d和 30d分别翅静脉采血 ,以常规血凝抑制试验检测抗体效价 ;用MTT法检测全血淋巴细胞对r MOMP的特异性增殖反应 ;免疫 30d使用鹦鹉热衣原体强毒株(BJF5株 )攻毒 ,观察临床症状 ,计算病变程度和保护率。结果表明免疫 14d后产生抗体 ,效价达 4 .2 (log2 ) ,免疫后 30d效价达到 5 .0 (log2 ) ;免疫组血液T淋巴细胞对r MOMP的增殖指数明显高于对照组 (P <0 .0 5 )。试验表明r MOMP在肉鸡体内可诱导对Cps特异性的体液免疫和细胞免疫应答 ,并能抵抗Cps强毒的攻击 ,显示r MOMP免疫肉鸡具有良好的免疫原性和保护性。     

Immune response after rabies vaccine in a kidney transplant recipient

A 48-year-old male kidney-transplant recipient was bitten by a rabid dog. His immunosuppressive treatment consisted of cyclosporine 60 mg b.i.d., mycophenolate mofetil (MMF) 250 mg t.i.d., and prednisone 5 mg. After wound care, he received 5 doses of purified vero cell rabies vaccine on days 0, 3, 7, 14, and 28, and human rabies immunoglobulin, according to international guidelines. Adequate levels of rabies virus neutralizing antibodies were observed after the administration of the third vaccine dose. However, a decrease of antibody titer was detected by day 28. Immunosuppressive medication was minimized, withdrawing MMF and reducing the dose of cyclosporine. Booster doses of the same vaccine were administered on days 38, 41, 45, 52, and 66. Adequate neutralizing antibody response was recovered during the ensuing 12 months, under reduced immunosuppression. Nineteen months after the incident, the patient remains with good graft function and is asymptomatic for rabies. It remains to be determined whether the attained immune response was either the result of the booster vaccinations or the reduction of immunosuppression alone. Nevertheless, such an outcome would have been possible only with the combined management strategy implemented.     

Oral Rabies Vaccination of Small Indian Mongooses (Urva auropunctata) with ONRAB via Ultralite Baits

The Ontario Rabies Vaccine (ONRAB) is a human adenovirus rabies glycoprotein recombinant oral vaccine immunogenic for small Indian mongooses when delivered by direct instillation into the oral cavity. We offered Ultralite baits containing ~1.8 mL 10^9.5 TCID₅₀ ONRAB oral rabies vaccine to 18 mongooses, while 6 mongooses were offered identical baits in placebo form. We collected sera from individual mongooses at days 0, 14 and 30 post vaccination (pv) and quantified rabies virus neutralizing antibodies (RVNA) using the rapid fluorescent focus inhibition test, with titers greater than or equal to 0.1 IU/mL considered positive. All study subjects were RVNA negative prior to bait offering. Bait consumption was variable: all 6 sham and 13 of 18 (72%) treatment animals consumed/punctured the baits offered. By day 30 pv, RVNA were detected among 11 of 13 (84.6%) of treatment mongooses that consumed/punctured baits, whereas sham-vaccinated mongooses remained RVNA negative throughout the study. We conclude ONRAB is immunogenic for mongooses by Ultralite bait delivery, although the bait design may need further optimization.     

Post-exposure prophylaxis with purified vero cell rabies vaccine during pregnancy--safety and immunogenicity

This study was conducted with the main objective of determining the safety and immunogenicity of purified vero cell rabies vaccine (PVRV) during pregnancy. Twenty nine pregnant women exposed to rabies were vaccinated with PVRV as per the Essen regimen advocated by World Health Organization. None of the women experienced any adverse side effects to the vaccine. The intrauterine growth and development monitored by ultrasound examination was found to be normal and the outcome of pregnancy was satisfactory. There were no congenital anomalies in any of the infants born and they were healthy and had normal growth and development during the one year follow-up period. The rabies neutralizing antibody titers from day 14 to day 365 following vaccination in these women was adequate and well above the minimum protective level of 0.5 iu/ml of serum. Protective levels of antibodies were also present in serum of some of the babies tested, for up to 3 months of age. The mothers and infants followed for one year period were doing well at the end of the study period. Consequently, PVRV was found safe and immunogenically efficacious during pregnancy.     

A critical evaluation of enzyme immunoassay kits for detection of antinuclear autoantibodies of defined specificities. II. Potential for quantitation of antibody content

OBJECTIVE: To analyze the performance of different commercial enzyme immunoassay (EIA) kits for measuring antibody levels of antinuclear antibodies (ANA) specific for double stranded (ds) DNA, SSB/La, Sm, and Scl-70. METHODS: Twenty companies that were known major purveyors of EIA kits for detection of ANA were approached to determine their interest and willingness to participate in this study. The manufacturers were advised that they would be sent coded sera containing mixtures of the Arthritis Foundation/Centers for Disease Control reference reagents, and that they were to use their own test kits to analyze the antibody specificities of these sera and to report the data, in optical density (OD) units, or their equivalent. The analysts were blinded to the concentration of the antibodies and the specificities. RESULTS: Initially, 11 manufacturers out of 20 agreed to participate, but 2 subsequently withdrew. The commercial EIA kits have the potential of being able to quantitate specific autoantibody content to ds-DNA, SSB/La, Sm, and Scl-70. However, certain deficiencies in these kits were also detected, the most obvious being lack of uniformly good performance, with kits of certain manufacturers showing exceptional accuracy in 3 out of 4 of their antibody-specific kits and poor accuracy for a 4th kit. CONCLUSION: It is important for clinicians to appreciate that there is marked inter-manufacturer variation in the performance of EIA kits used as an aid in the diagnosis of systemic rheumatic diseases. Manufacturers need to exercise constant surveillance of kit performance and to provide assurance that such is being done. Improved EIA kits would lend themselves to reliable quantitation of antibody levels in human sera and help to determine whether serial measurement of antibody levels might be useful in monitoring disease activity.     

Antibody response after a four-site intradermal booster vaccination with cell-culture rabies vaccine.

The current World Health Organization recommendation for booster vaccination of previously immunized individuals with potential exposure to rabies is two doses of vaccine intramuscularly or intradermally on days 0 and 3. We report responses to two types of postexposure treatment of healthy individuals who had received preexposure rabies vaccination 1 year previously. Group A individuals received four intradermal doses (one-fifth of the diluent volume of vaccine per dose) on day 0, and group B individuals received two intramuscular doses on days 0 and 3. Immunogenicity of the two booster regimens was assessed by titrating the amount of neutralizing antibody (Nab). We found that the booster doses of vaccine produced remarkable responses in all subjects. Nab titers of > or = 0.5 IU/mL (acceptable antibody level for protection against rabies) were detected in all subjects on day 14, and they were shown to be consistently high 1 year after the booster vaccination. We also found that the Nab titers for group A were significantly higher (two- to eightfold) than those for group B on days 5, 14, 150, and 360 after the initial booster vaccination (P < .05). Our study shows that the four-site intradermal booster regimen with use of one-fifth of the diluent volume of cell-culture rabies vaccine on day 0 is associated with a significantly higher antibody response than is the conventional booster regimen for subsequent postexposure rabies treatment of individuals who have received preexposure rabies vaccination with cell-culture rabies vaccine 1 year previously.