布鲁氏菌特异性抗原BSAg—ELISA试验检测病人血清抗体的研究

以布鲁氏菌特异性抗原ＢＳＡｇ进行ＥＬＩＳＡ试验，检查经常规血清学试验确诊的布鲁氏菌病人血清１５５份，全部阳性，阳性率为１００％。检查常规实验室视为疑似布鲁氏菌病人血清８０份，ＥＬＩＳＡ试验阳性２４份，阳性率３０％，检查布鲁氏菌病疫区内健康人血清１００份、ＥＬＩＳＡ试验均为阴性，但有１２份血清ＥＬＩＳＡ滴度达１：１００。因此认为ＢＳＡｇ－ＥＬＩＳＡ试验特异性强，敏感性高，可用于布鲁氏菌病血清学诊断。     

布鲁氏菌特异性抗原BSAg─ELISA试验检测病人血清抗体的研究

以布鲁氏菌特异性抗原ＢＳＡｇ进行ＥＬＩＳＡ试验，检查经常规血清学试验确诊的布鲁氏菌病人血清１５５份（其中急性５１份，慢性１０４份），全部阳性（滴度≥１：４００），阳性率为１００％。检查常规实验室视为疑似布鲁氏菌病人血清８０份，ＥＬＩＳＡ试验阳性２４份，阳性率３０％，检查布鲁氏菌病疫区内健康人血清１００份、ＥＬＩＳＡ试验均为阴性，但有１２份血清ＥＬＩＳＡ滴度达１：１００。因此认为ＢＳＡｇ─ＥＬＩＳＡ试验特异性强，敏感性高，可用于布鲁氏菌病血清学诊断。     

流行性出血热的血清学研究

为了探索流行性出血热的诊断方法,作者曾先后作过血清沉淀反应、补体结合试验、梅毒血清反应和外一裴氏反应,以上试验因阳性率低,或缺乏特异性,故诊断价值不大。作者曾对7名典型的流行性出血热患者和6名恢复期患者作皮内试验,其结果100%阳性。而用同样方法对斑疹伤寒、猩     

滴金免疫渗滤法快速诊断肺吸虫病的研究

目的寻找一种快速诊断肺吸虫病的方法.方法以肺吸虫成虫浸出液为抗原,金标抗人IgG为显示剂检测肺吸虫抗体(DIGFA).结果检测肺吸虫病人血清41份,阳性符合率为100%;检测非流行区健康人血清51份,流行区健康人血清30份,阴性符合率100%;检测班氏丝虫病人血清20份,均为阴性;检测日本血吸虫病人血清20份,其中2例呈阳性.用DIGFA与ELISA同时检测肺吸虫病人血清41份,非流行区健康人血清51份及流行区健康人血清30份,两种方法的阳性与阴性符合率均为100%.结论DIGFA检测人体肺吸虫抗体敏感性高,特异性强,检测时无需特定的仪器,操作简便、快速,适于基层单位和现场应用.     

人体丝虫病对异种丝虫抗原的免疫反应

作者用犬丝虫(Diro)、棉鼠丝虫(Lito)及血管国线虫Angiostrongylus cantonensis(Ac)等3种抗原对冲绳地区98份血清,其中15份系班氏丝虫微丝蚴阳性者,83份系微丝蚴阴性者;又对塔希提岛18例班氏丝虫患者进行了间接红细胞凝集(HA)试验、补体结合(CF)试验和琼脂扩散(ID)试验。此外对实验感染棉鼠丝虫的棉鼠血清与异种丝     

胶体染料斑点兔疫试验法(DIA)检测血吸虫病人血清抗体的进一步研究

本研究对胶体染料斑点免疫试验法(DIA)检测血吸虫病人抗体的有关因素进行了系统的观察,并对实验方法进行了改进.用改进后的DIA检测288份血吸虫病人血清抗体,其中69份急性病人血清,阳性率为95.7%;110份慢性病人血清阳性率为96.4%;19份晚期病人血清阳性率为47.4%;40份华支睾吸虫病人血清,2份阳性,交叉反应率为5%;50份健康人血清,1份阳性,假阳性率2%.敏感性、特异性与酶联斑点免疫试验法相似.改进后的DLA简单、经济、易行,适于现场应用.     

斑点免疫金渗滤法检测弓形虫感染效果

目的建立一种简捷、准确的弓形虫感染检测方法。方法将弓形虫P30抗原包被在硝酸纤维素膜上,与待检血清中的相应抗体结合,通过金标记Protein-A蛋白直接显色。结果用该试剂检测5种血清样本共221份,其中弓形虫感染病人血清58份,敏感性为96.55%(56/58)。正常人群对照血清68份,检测结果全部为阴性,特异性为100%。检测25份类风湿病人血清、18份血吸虫病人血清及22份肺炎支原体病人血清,仅有1例类风湿病人血清出现阳性交叉反应。结论用弓形虫P30为抗原,以金标记Protein-A蛋白为探针显色系统的斑点免疫金渗滤法,对检测弓形虫病人血清抗体具有较高的敏感性和特异性,且操作简单、快速。     

一种改良的布鲁氏菌病抗体微量凝集检测方法的建立与评价

目的 建立一种微量的布鲁氏菌病抗体凝集检测方法,用于布鲁氏菌病高通量检测。方法 依据我国《布鲁氏菌病诊断标准》(WS269-2007)规定的病人诊断标准,用试管凝集试验做诊断标准。通过优化微量凝集试验抗原浓度,对142份疑似病人血清同时做试管凝集和微量凝集试验,进行效果评价。结果 最优的微量凝集抗原浓度为1:10,建立微量凝集法具有较高的敏感度和特异度,分别为98.9%和92.3%;和常规试管凝集法相比,两种方法符合率为96.5%。常规试管凝集检测得到21份阴性样本,22份可疑样本(1:50),99份阳性样本(>1:100)。微量凝集检测得到30份阴性样本、17份可疑样本(1:50)、95份阳性样本(>1:100)。两种试验方法,结果相同的占70.4%(100/142),经统计学检验差异无统计学意义(χ²=0.8,P>0.05)。结论 建立一种可显色的布鲁氏菌病抗体微量凝集检测方法,可以在96孔V型板上同时检测24份标本,且结果易判读,更适宜基层防疫人员现场检测,最终为疫情处置提供强有力的技术支持。     

免疫酶染色试验检测斯氏狸殖吸虫抗体的研究

目的探索用免疫酶染色试验检测斯氏狸殖吸虫病抗体的敏感性和特异性。方法从病犬肺虫囊中收集斯氏狸殖吸虫成虫,冰冻切片,制作抗原片。采用免疫酶染色试验(IEST)检测斯氏狸殖吸虫病人和病鼠血清抗体。试验设健康大鼠血清对照及其他寄生虫病血清对照。结果斯氏狸殖吸虫病人和病鼠血清特异抗体阳性率分别为96.7%和97.4%。实验动物从感染后第2周开始抗体检测阳性,第4周阳性率达97.4%,持续10周。结论免疫酶染色试验敏感性高、特异性强,对斯氏狸殖吸虫病的早期诊断有重要参考价值。     

血吸虫病补体结合抗原的比较评价

许多研究者认为补体结合试验是最好的血吸虫病血清诊断方法。但由于病人感染度、感染时间、年龄及血吸虫种等因素的差异,要比较不同实验室所得的结果是困难的。本文的目的是用同批血清比较Fairley氏螺“肝”抗原与其它多种抗原的效价。血清来自:(1)31名未治疗的曼氏血吸虫     

Influence of brucellosis history on serological diagnosis and evolution of patients with acute brucellosis

Serological diagnosis of human brucellosis is problematic in endemic brucellosis regions and with patients having a history of brucellosis. The aim of this study is to ascertain the serologic and evolutionary behavior of the tests of serum agglutination, Coombs anti-Brucella, immunocapture-agglutination, enzyme-linked immunosorbent assay (ELISA) IgG, IgA, IgM and ELISA-IgG avidity against Brucella lipopolysaccharide (S-LPS), in patients with acute brucellosis based on whether or not a history of brucellosis exists. Titers and seropositivity in all the tests assayed were higher in the patients having brucellosis history (from 90.9% in ELISA-IgM to 100% in ELISA-IgG) than in the patients lacking such history (from 79.3% in ELISA-IgM to 86.2% in Coombs, immunocapture-agglutination, and ELISA-IgG). IgG S-LPS avidity results in patients with brucellosis history were significantly higher (always over 84%) than in patients without brucellosis history (from 48.0% in the initial sera to 81% ten months later) (p<0.001). The titers of antibodies against Brucella in the initial sera and ELISA-IgG avidity against S-LPS may allow distinguishing patients with brucellosis caused by primary infection in the initial stages of the disease from patients seropositive due to prior infections from Brucella.     

量子点标记免疫层析技术检测布鲁氏菌病的方法研究

目的 通过研究量子点标记免疫层析技术检测布鲁氏菌病抗体,制备量子点免疫层析试纸条,验证快速检测布鲁氏菌病的可行性。方法 将蛋白A和布鲁氏菌全菌蛋白分别作为质控线和检测线喷涂在硝酸纤维素膜上,以量子点标记布鲁氏菌全菌蛋白,稀释后喷涂在玻璃纤维素膜上,最后进行组装、切割制成检测用试纸条。应用该试纸条检测布鲁氏菌病患者样本102例,健康人血清47例,以试管凝集试验为对照,比较两组方法的符合率。结果 建立的量子点免疫层析试纸条性能较好,布鲁氏菌病抗体的检测敏感性为99.0%,特异度为95.8%,两者符合率为98.0%;将阳性血清稀释至128倍,通过荧光检测仪仍可检测到T线荧光值,该纸条重复性好,储存时间30 d内稳定性较好,其变异系数为4.1%。结论 该方法操作简单、快速稳定、灵敏度高、成本低,15 min内完成检测,血清用量低,可实现现场快速检测,在布鲁氏菌病的早期检测中具有良好前景。     

Changes in IgM and IgG antibody concentrations in brucellosis over time: importance for diagnosis and follow-up

Changes in concentrations of IgM and IgG antibodies to Brucella were monitored for at least 13 mo by an enzyme-linked immunosorbent assay (ELISA) in 52 patients with culture-positive brucellosis. Two main patterns were observed. After an initial peak, 29 patients (56%) had a steady drop in their IgG levels, whereas 23 (44%) had more than one peak over time. All patients with a chronic form of brucellosis or a relapse were in the second group. In most cases, Brucella antibodies, although falling to low levels, remained measurable. Cutoff levels for IgM and IgG were calculated after considering serum antibody concentrations in people who had recovered from an infection. A separate normal range was established for occupationally exposed workers. On admission, sera from all patients contained Brucella antibody levels greater than established cutoff levels. Our results show that ELISA is an excellent method for diagnosis and follow-up of brucellosis.     

Seroepidemiological study on human brucellosis in Assiut Governorate

Brucellosis is the most important zoonotic disease constituting a public health problem in Assiut Governorate, hence this study was carried out to determine the prevalence of brucellosis among humans in Assiut Governorate. A total of 7154 peripheral blood samples were collected from patients with fever at Assiut Fever Hospital during the period from 2002-2003. A full detailed anamnestic and clinical assessment in the form of questionnaire was designed for each individual to determine the risk factors with specific emphasis to age, sex, residence and occupation. All serum samples were screened for Brucella antibodies by slide agglutination test. Positive sera were further analyzed by standared tube agglutination test. Enzyme linked immunosorbent assay (ELISA) was carried out to detect IgM and IgG Brucella antibodies. Statistical analysis was performed and correlation coefficient was done between all risk factors. Results declared that the prevalence of brucellosis was (1.29 +/- 0.004 %) and (1.22 +/- 0.002 %) as detected by agglutination and ELISA, respectively. IgM antibodies were estimated in 9.8 % of the examined patients, while IgG antibodies were found in 30.4 % of the examined patients, moreover both IgM and IgG antibodies were detected in 54.3 % of the examined patients. The prevalence of brucellosis was significantly (P < 0.05) affected by sex, where the rate of detection was higher among females (1.76 +/- 0.009 %) than males (1.05 +/- 0.004 %) as detected by agglutination test. On the other hand, the prevalence rate based on ELISA was (1.64 +/- 0.39 % and 1.01 +/- 0.89 %) for females and males, respectively. Prevalence of brucellosis was higher in rural areas (1.3 +/- 0.005 % & 1.25 +/- 0.009 %) than in urban areas (1.23 +/- 0.001% & 1.12 +/- 0.01 %) as detected by agglutination test and ELISA, respectively. The prevalence of brucellosis varied significanctly between different occupational and age groups. Public health impact of brucellosis is discussed and suggestive measures for control are explained.     

双抗原夹心酶联免疫试验检测急性布鲁杆菌病患者抗体真实性观察

目的观察双抗原夹心酶联免疫试验(DAgS-ELISA)检测急性布鲁杆菌病(布病)患者的真实性,即灵敏度和特异度。方法以试管凝集试验(SAT)法为金标准,用DAgS-ELISA与虎红平板凝集试验(RBPT)、补体结合试验(CFT)以及间接酶联免疫试验(I-ELISA)对同一布病患者人群和对照组人群血清标本进行检测比较。结果真实性比较,与金标准SAT的符合率分别为:DAgS-ELISA 96.49%,RBPT 94.74%,I-ELISA 85.09%,CFT 72.81%;诊断效能比较,阳性和阴性预测值DAgS-ELISA分别为93.48%、98.53%,RBPT分别为88.00%、100%,CFT分别为100%、69.30%,I-ELISA分别为78.72%、89.55%。结论DAgS-ELISA检测急性布病患者的敏感性和特异性优于其他方法。     

抗生物素蛋白-生物素酶标法在检测布鲁氏菌病抗体中的应用

本文报道使用抗生物素蛋白—生物素酶标法(简称ABC-ELISA法)检测布氏菌抗体,以提高对布病患者的检出率。用ABC-ELISA等五种方法检测了100名健康人及150名慢性布病患者的血清,结果表明ABC-ELISA法检测布氏菌病抗体有较高的敏感性,其血清滴度比2-MET高64倍,比Wright氏反应高8—32倍,比Coomb’s试验高4—16倍,比PPA-ELISA高4—8倍。其检出率明显高于其他四项试验(P<0.01),并有较强的特异性,是布病诊断中有发展前途的新方法。     

布鲁氏菌病快速检测试纸条的探索

目的 建立一种操作简单、结果肉眼可视,并具有良好敏感性和特异性的适合现场快速检测布鲁氏菌病的胶体金层析方法。方法 将重组的布鲁氏菌外膜蛋白OMP22和OMP28作为胶体金标记物,抗OMP22单克隆抗体作为质控线,OMP22和OMP28蛋白作为检测线组装试纸条。结果 该试纸条检测牛布鲁氏菌标准阳性血清最大稀释度可达1:128;并且与牛结核、蓝舌病、牛病毒性腹泻、口蹄疫、牛白血病及小反刍兽疫血清无交叉反应;检测牛、羊源血清样品结果与商品化的ELISA检测试剂盒(IDEXX)符合率为95%,与虎红平板凝集试验(RBPT)检测的符合率为97.4%。结论 胶体金层析试纸条在检测布鲁氏菌病方面具有快速、敏感性高及特异性强的特点,适用于布鲁氏菌病动态流行病学调查和临床快速筛查。     

2001-2015年锦州市布鲁氏菌病疫情流行病学分析

目的描述2001-2015年锦州市布病疫情流行病学特征和变化趋势,探讨下一步的防控重点。方法用描述流行病学方法对锦州市布病疫情进行分析,对试管凝集试验阳性率与发病率进行Pearson相关性分析。结果 2001-2015年锦州市布病发病率为0.45/10万-16.53/10万。发病时间集中在3-7月,占总数的66.89%。2003-2015年共采集重点人群血清5 904份,阳性率为5.01%。2007-2015年从136份血培养标本中分离出布鲁氏菌63株,其中羊种3型48株、羊种1型12株,羊种变异2株、犬种1株。结论锦州市近年布病疫情呈明显上升趋势,其优势菌株主要是羊种3型。对犬的布鲁氏菌携带情况应予以一定的关注,及时淘汰病犬。     

斑点金免疫渗滤法快速诊断布鲁杆菌病的研究

目的建立一种布鲁杆菌病快速免疫诊断方法。方法以硝酸纤维素膜为载体,胶体金标记蛋白为显示剂,通过最佳实验条件的选择及稳定性、重现性、敏感性、特异性和交叉反应试验,建立斑点金免疫渗滤法(DIGFA)快速检测布鲁杆菌病。结果 DIGFA快速检测布鲁杆菌病具有很好的重现性和稳定性;敏感性、特异性分别达到98％和100％;未发现与肺结核患者、乙肝患者及幽门螺杆菌感染者血清有交叉反应。结论 DIGFA快速检测布鲁杆菌病快速、简便,测试过程1-2 min内完成,试剂敏感、特异、稳定,非常适合布鲁杆菌病的诊断及流行病学调查。     

滴金免疫测定法检测不同人群布鲁氏菌病

目的探讨滴金免疫测定法在不同人群中检测布鲁氏菌感染的应用价值。方法用已建立的布病滴金免疫测定技术检测不同职业和不同感染类型人员的布病抗体,并与布病试管凝集试验作平行检测。结果滴金免疫测定法与试管凝集试验两种方法符合率为99.88%,阳性符合率达到94.25%;且本法对布病发病人群和高滴度血清样本具有更高的检测敏感性;对不同职业人群检测,接触山羊和奶牛职业人群的布鲁氏菌病抗体检出率分别为6.40%和1.32%。结论滴金免疫测定法对布病疫区人群和非疫区重点职业人员开展布病监测具有实用价值,也可用于布病的临床检测和流行病学筛查。