布病特异性血清学检测技术应用概述

布病是由布鲁菌属细菌侵入机体引起的传染-变态反应性疾病。目前,该病在世界范围内广泛流行,主要侵犯人类、家畜及其它多种动物,部分人或动物感染后并无明显临床表现,不能为临床布病确诊提供可靠依据。因此,准确、快速的检测诊断是预防和控制布病的关键环节。本文就目前国内外用于布病检测诊断的方法平板凝集试验(PAT)、琥红平板凝集试验(RBPT)、试管凝集试验(SAT)、补体结合试验(CFT)、酶联免疫吸附试验(ELISA)、荧光偏振试验(FPA) 、免疫胶体金技术(GICA)等特异性血清学检测技术予以概述。     

建立双抗原夹心酶免疫试验对人畜布鲁氏菌抗体检测的研究

目的为人畜布鲁氏菌病(布病)的血清学诊断、监测及流行病学调查提供特异、敏感、快速的试验方法。方法在制备高滴度的布鲁氏菌(布氏菌)抗原及其抗原辣根过氧化物酶结合物基础上。建立了检测人畜布氏菌抗体的双抗原夹心酶免疫试验(DAgS-EIA),包括常规双抗原夹心酶联免疫吸附试验(DAgS-ELISA)、双抗原夹心酶免疫斑点试验(DAgS-DIEA)以及快速双抗原夹心ELISA(快速DAgS-ELISA),并对影响本试验的主要因素因素进行了试验。结果制备了高滴度的布氏菌抗原及其抗原酶结合物和冻干制品。并在此基础上,建立了常规DAgS-ELISA及DAgS-DIEA以及快速DAgS-ELISA检测人畜布氏菌抗体方法。经对115份布病患者血清检测结果,阳性率以DAgS-ELISA为最高(61.7%),其余依次为RBPT(58.1%)I、-ELISA(55.6%)、DAgS-DIEA(53.7%)、及SAT(44.2%)且用DAgS-EIA检测布氏菌感染羊、牛、猪及实验动物家兔、豚鼠和小鼠均为强阳性反应,而对正常人、羊、牛、猪及实验动物血清均为阴性反应。结论双抗原夹心酶免疫试验检测人畜血清布氏菌抗体,不仅特异、敏感、快速、简便而且仅需制备单一的布氏菌抗原酶结合物就可对人及各类动物血清布氏菌抗体进行检测。     

不同血清抗体检测方法在布病监测中的意义

以往对人群进行布病流行病学监测时,最常采用的血清抗体检测方法有玻片凝集试验(PAT)、试管凝集试验(SAT)、二疏基乙醇凝集试验(2—ME)、补体结合试验(CFT)和Coomb′s试验,并以此来辨别与布病有关的各类特异性抗体。     

用布氏菌乳环抗原的5种血清学试验检测人畜布氏菌抗体的研究

目的摸索建立用布氏菌乳环抗原的血清学试验,以用于人畜血清中布氏菌抗体的检测。方法在制备并标化布氏菌乳环抗原的基础上,用此抗原分别做PAT、SAT、CAT、CFT、及DAgS-ELISA,同时与这些试验的常规方法对部分布氏菌感染的人畜血清及正常血清进行对比检测。结果5种常规血清学试验即RBPT、SAT、CAT、CFT及DAgS-ELISA与用布氏菌乳环抗原的5种血清学试验即PAT、SAT、CAT、CFT及DAgS-ELISA对部分布氏菌感染人畜血清进行对比检测,其阳性率分别为91.5%(65/71)、83.3%(60/72)、58.2%(39/67)、46.4%(26/56)、81.9%(59/72)以及94.4%(67/71)、83.3%(60/72)、56.7%(38/67)、54.5%(30/55)、80.6%(58/72)。并且对两种抗原的试验所得阳性率进行显著性差异检验,差异均无显著性。(χ2分别为0.220、0.000、0.950、0.360、0.005,P>0.05)。其所测抗体的几何平均滴度(GMT)分别为1∶121、1∶23、1∶6和1∶19以及1∶109、1∶23、1∶10和1∶19。结论用布氏菌乳环抗原不仅适用做MRT对布氏菌感染奶特异性抗体的检测,而且可用此种抗原PAT、SAT、CAT、CFT及DAgS-ELISA对人畜血清布氏菌抗体的检测。     

浙江省台州地区家畜布鲁氏菌病血清学、细菌学诊断

<正> 应用普通平板凝集试验(RAT)、虎红平板凝集试验(RBPT)、试管凝集试验(SAT)、半胱氨酸凝集试验(CAT)和补体结合试验(CFT)等五种血清学试验方法检查我区家畜布鲁氏菌病20887头,查出130头阳性,阳性率为0.067％,故认定我区“布病”存在。但没有分离到布氏菌(浙江省未分离到)     

酶联免疫吸附双抗原夹心法检测人畜布氏菌抗体的研究

目的 为人畜布氏菌病血清学诊断、监测及流行病学调查提供实用和简便的一种酶免疫试验技术。方法 在制备高滴度布氏菌抗原辣根过氧化物酶结合物及其冻干制品基础上，应用酶联免疫吸附双抗原夹心法即双抗原夹心酶联免疫吸附试验（DAgS－ELISA）、试管凝集试验（SAT）及虎红平板凝集试验（RBPT）对从山西省布氏菌病疫区收集的布氏菌病患、布氏菌感染羊、牛、猪以及健康人、羊、牛、猪共计290份血清进行对比检测。结果 上述3种试验对健康人、羊、牛、猪共计140份血清检查均为阴性反应，用DAgS-ELISA对布氏菌感染人、羊、牛、猪共计150份血清检查，其阳性率分别为91．4％、74．6％、66．7％及66．7％，从总体而言，检查的特异性及敏感性，以DAgS-ELISA优于SAT及RBPT。结论 双抗原夹心酶联免疫吸附试验检测人畜布氏菌抗体，不仅特异、敏感、简便，而且制备一种布氏菌抗原酶结合物及其冻干制品，可用于人及各种动物布氏菌抗体的检测，值得推广应用。     

鞍山市2015-2020年布鲁菌病血清学检测分析

目的 通过分析布鲁菌病血清学检测结果,了解布病的发病情况,预测流行趋势,为制定防治对策提供依据.方法 对鞍山市门诊接诊的布病疑似患者进行血清学检测,按照《布鲁氏菌病诊断标准》执行,采用虎红平板凝集法(RBPT)初筛试验,结果阳性者,采用试管凝集试验(SAT)作为确认试验.结果 共检测布病可疑患者815例,阳性率61.9...     

浅析温度和时间对RBPT试验的影响

目前,对布氏菌病(简称布病)的临床检验主要有2种,即 RBPT(虎红平板凝集试验)和 SAT(特莱氏试验或试管凝集试验)。虽然 RBPT 具有方法简便、快捷,易于操作和结果便于观察且漏检率极低的优点,但易出现假阳性,假阳性率较高,一般只作为布病的大面积筛查,在临床上需要借助 SAT 来确认。如何提高 RBPT 阳性率,怎样排除或减少 RBPT假阳性,我们在近20年的布病临床检验中,发现温度和时间对 RBPT 试验的影响很大。近10年来,我们对部分疑似布病患者的血清,经过不同时间和不同温度     

2008年宁夏中卫市沙坡头区人间布鲁杆菌病调查与分析

目的 调查宁夏中卫市沙坡头区布鲁杆菌病(简称布病)的流行现状及特征.方法 2008年采取分层随机抽样的方法,对宁夏中卫市沙坡头区布病重点人群进行流行病学调查,应用琥红平板凝聚试验(RBPT)和试管凝集试验(SAT)进行血清学检测,按照<布鲁氏菌病诊断标准及处理原则>(GB 15988-1995)诊断病例.结果 共调查10个乡镇布病重点人群2480人,检测血清604人份,布病感染者28例,感染率为4.64%(28/604).检出布病患者15例,均为奶牛饲养人员,检出率为2.48%(15/604);饲养人员感染率最高,为6.45%(26/403).结论 宁夏中卫市沙坡头区存在人间布病的流行,患病与职业有关.     

2008年宁夏中卫市沙坡头区人间布鲁杆菌病调查与分析

目的 调查宁夏中卫市沙坡头区布鲁杆菌病(简称布病)的流行现状及特征.方法 2008年采取分层随机抽样的方法,对宁夏中卫市沙坡头区布病重点人群进行流行病学调查,应用琥红平板凝聚试验(RBPT)和试管凝集试验(SAT)进行血清学检测,按照<布鲁氏菌病诊断标准及处理原则>(GB 15988-1995)诊断病例.结果 共调查10个乡镇布病重点人群2480人,检测血清604人份,布病感染者28例,感染率为4.64%(28/604).检出布病患者15例,均为奶牛饲养人员,检出率为2.48%(15/604);饲养人员感染率最高,为6.45%(26/403).结论 宁夏中卫市沙坡头区存在人间布病的流行,患病与职业有关.     

胶体金免疫层析技术在羊布鲁氏菌病的检测应用

目的 了解胶体金免疫层析技术(GICA)检测布鲁氏菌病血清的效果。方法 选择格尔木送检的299份羊血清,分别采用GICA、虎红试管凝集试验(RBPT)和试管凝集试验(SAT)3种方法进行布病检测,分析3种方法检测结果的一致性。结果 3种方法的检测结果进行两两对比后,RBPT与 SAT的一致性检验结果kappa值为0.972(χ²=282.563,P<0.001);GICA与SAT的一致性检验结果kappa值为0.940(χ²=256.577,P<0.001)。结论 说明GICA的实验结果与SAT的实验结果具有高度一致性。     

酶标金葡菌A蛋白结合物用于猪种布病疫区人群血清流行病学检查的价值

本文介绍了应用HRP-SPA结合物,作为第二抗体做ELISA试验,用于猪种布病疫区人群血清流行病学检查。本方法重复性好,特异性强,敏感性高。用该法对猪种布病疫区1168份人血清进行检查,同时用布病血检常规(SAT、CFT、PAT、RBPT、AGT),进行对比检定,结果表明:无论对人猪种布病的诊断或疫区人群的普查,ELISA的阳性率都比SAT、CFT、RBPT、PAT、AGT高。ELISA与SAT的符合率为92.6％。发现ELISA与SAT之间有良好的正相关;HRP-SPA结合物与HRP-兔抗人IgG结合物敏感性、特异性相似。认为利用HRP-SPA作为第二抗体的ELISA技术,无论对人猪种布病的诊断或疫区人群进行大规模血清流行病学检查都是适用的,有推广的价值。     

青藏高原喜马拉雅旱獭布鲁氏菌病血清流行病学调查分析

目的 检测青海省喜马拉雅旱獭布鲁氏菌抗体,确定青海喜马拉雅旱獭布鲁氏菌病的地理分布.方法 采用描述性流行病学研究中的现况调查方法,选取青海省人、畜布鲁氏菌病流行地区,有喜马拉雅旱獭栖息的部分县(市),采集青藏高原喜马拉雅旱獭血样,采用胶体金免疫层析技术(GICA)、虎红平板凝集试验(RBPT)、试管凝集试验(SA T)...     

天津市农牧场系统布鲁氏菌病流行情况的初步调查

本文对天津部分奶牛队643名职工,进行了临床和 HT、SAT、CFT、RBPT、ELISA 等血清学检测和皮内变态反应试验。查得血清阳性率7.93%(51/643),患病率为2.33%(15/643)。并且对阳性者分别按性别、年龄、接触奶牛年限、职业(工种)、血型等进行了分析。得出了感染与年龄、接触年限、职业(工种)、血型等在统计学上显著有关。指出:患布鲁氏菌病的奶牛是人畜间布鲁氏菌病流行的传染源,并指出以接触为主的传播途径和牛种布鲁氏菌为流行菌株。最后强调对重点人群布鲁氏菌病普查的重要性,并对今后的布鲁氏菌病防治工作提出了建议。     

Evaluation of serological tests for diagnosis of brucellosis

The aim of the present study was to compare serological tests (Rose Bengal [RB]; standard agglutination test [SAT]; enzyme immunoassay [EIA] for detection of IgM, IgA, and IgG; and 2-mercaptoethanol [2-ME] test) that are routinely used in patients prediagnosed with different clinical types of brucellosis (acute, subacute, or chronic), and to evaluate the results of the IgG avidity test. Ninety-two patients having titers≥1/160 as measured by SAT were included in the study. The IgG avidity test was performed in 78 patients who had positive EIA-IgG results. RB test results were positive in 88 (95.7%) patients. A statistically significant correlation was found between a positive EIA-IgM result and the diagnosis of acute brucellosis. When compared to the results of the SAT, the 2-ME test showed a lower titer in 55 (59.8%) patients, and the agreement between the 2-ME test and EIA-IgG was calculated as 84.8%. No statistical difference was found between the 40% avidity index used in the IgG avidity test and avidity maturation time (6 months). From our study, we concluded that (i) the RB and SAT tests are appropriate and reliable tests for the serological diagnosis of brucellosis; (ii) IgM can be used as a marker of acute brucellosis; (iii) the 2-ME test, similar to EIA, can be used to determine IgM levels; and (iv) the IgG avidity test should be standardized.     

2009年陕西省渭南市人间布鲁杆菌病监测结果分析

目的 了解陕西省渭南市布鲁杆菌病(简称布病)流行状况,为制订防治策略提供科学依据.方法 根据<陕西省布鲁氏菌病监测工作实施方案>,2009年在渭南市选择澄城、大荔、合阳、潼关、韩城5个县(市)的固定监测点35个乡(镇)、非固定监测点24个乡(镇)开展人间布病监测工作,对7～60岁与家畜及畜产品有接触的重点人群进行临床检查,有症状者进行血清学检测(虎红平板凝集试验和试管凝集试验).操作方法及判定标准均按<布鲁氏菌病诊断标准>(WS 269-2007)进行.结果 固定监测点共调查重点人群8664人,做虎红平板凝集试验1407人,阳性27人,阳性率为1.92%(27/1407);做试管凝集试验27人,阳性27人,阳性率为100%(27/27);确诊25例,确诊率为92.59%(25/27).非同定监测点共调查重点人群3464人,做虎红平板凝集试验411人,阳性3人,阳性率为0.73%(3/411);做试管凝集试验3人,均为阳性,且全部确诊.共确诊新发病例28例,人间布病感染率为0.25%(30/12 128),发病率为2.06/10万(28/1361618).结论 渭南市人间布病疫情居高不下,建议政府加大经费投入,积极开展畜间和人间布病疫情监测工作,控制疫情扩大蔓延.

Abstract：

Objective To survey and analyze characteristics of brucellosis epidemic in Weinan city of Shaanxi province for the purpose of setting up prevention and control measures for the disease. Methods According to "The Executing Plan for the Work of Surveying Brucellosis Disease in Shaanxi Province", 35 villages(towns) of designated monitoring locations and 24 villages (towns) of randomized monitoring locations in five countries of Weinan were chosen to survey brucellosis disease. The five countries were Chengcheng, Dali, Heyang, Tongguan and Hancheng. High risk populations with a history of contacting livestock and livestock products aged between 7 and 60 underwent clinical and serology examination[rose bengal plate agglutination test(RBPT) and standard tube agglutination test(SAT)]. All manipulation methods and judging standards were in accord with the "Diagnostic Standard for Brucellosis" (WS 269-2007). Results In the designated monitoring location, a total of 8664 people at high risk were investigated, among whom 1407 people were tested by RBPT test and 27 people were positive,the positive rate was 1.92%(27/1407); 27 people were tested by SAT test and 27 people were positive, the positive rate was 100% (27/27); 25 people were diagnosed and the diagnosis rate was 92.59%(25/27). In the randomized monitoring location, a total of 3464 people at high risk were investigated, among whom 411 people were tested by RBPT test and 3 people were positive, the positive rate was 0.73%(3/411 ), 3 people were tested by SAT test which were all positive and made a definite diagnosis. Twenty-eight new cases were made a definite diagnosis and its incidence was 2.06 in a hundred thousand(28/1 361 618). Conclusions The infection of human brucellosis in Weinan city stays at higher level. The governments should increase input for the monitoring,investigating and disinfecting to prevent the disease from increasing and outspreading.     

2006-2010年青海省人间布鲁杆菌病调查分析

目的 分析青海省布鲁杆菌病(简称布病)的流行特征及其影响因素,为制订布病防制对策提供科学依据.方法 2006- 2010年,选择青海省布病高发地区以及青海省中央补助地方公共卫生专项资金人间布病预防控制项目的5个县(河南县、达日县、天峻县、平安县、海晏县)为调查点,同时对青海省生物药品厂高危从业人员进行调查.对调查对象进行布鲁杆菌素皮内变态反应试验,同时按知情同意原则,采集血清,做虎红平板凝集试验(RBPT)和试管凝集试验(SAT).按《全国人间布鲁氏菌病监测方案(试行)》中的调查表进行流行病学调查,询问流行病学接触史,结合临床症状和体征,按照《布鲁氏菌病诊断标准及处理原则》(GB 15988-1995)和《布鲁氏菌病诊断标准》(WS 269-2007)进行检测确诊病人.结果 在被检测的8368份血清样品中,RBPT检出阳性347份,阳性率为4.15％;SAT检测血样5346份,阳性180份,阳性率为3.37％.2009年6月对青海省生物制药厂进行跟踪调查,全厂共调查112人份,RBPT检出阳性83份,阳性率为74.11％,SAT检出阳性58份,阳性率为51.79％,确定布病新发病人8例,布病慢性期病人4例.2006 - 2010年青海省报告新发布病25例,发病高峰为每年的3-7月份,病例以牧民为主.结论 加强动物检疫,强化宣传教育、提高防护意识,可切实控制布病疫情.     

Evaluation of Brucellacapt for the diagnosis of human brucellosis

OBJECTIVE: To evaluate the role of Brucellacapt in the diagnosis of human brucellosis, and the correlation with the evolution of the disease. METHODS: Twenty-six patients who were admitted to the General Hospital of Albacete (Spain) over a 2-year period and diagnosed with brucellosis were included in the study. One hundred and twenty-three serum samples collected at the time of diagnosis and at intervals during and after treatment were tested by the Coombs test, the standard seroagglutination test (SAT), and Brucellacapt (a new test based on an immunocapture-agglutination technique). To study the specificity of Brucellacapt, sera from 20 patients with other infectious diseases and 20 sera from healthy donors were included in this study. RESULTS: The sensitivity of the Brucellacapt at the moment of diagnosis was similar to the Coombs test (96 and 100%, respectively), somewhat higher than that of SAT (73%). And the specificity of the Brucellacapt (97.5%) was less than SAT and the Coombs test, that was 100%. The correlation between the classical serological tests and Brucellacapt, showed that titers in Brucellacapt and Coombs test of patients were both similar in a range of 1-2 dilutions. The correlation between Brucellacapt and Coombs (r=0.14) and between Brucellacapt and SAT (r=0.0) did not reach statistical significance. However, the correlation coefficient between Coombs and SAT was r=0.8. CONCLUSIONS: Brucellacapt and Coombs tests showed a similar sensitivity and specificity in the diagnosis of human brucellosis. In addition, as Coombs test, Brucellacapt could help to diagnose patients with long evolution of brucellosis that are not detected with SAT.     

Limitations of the standard agglutination test for detecting patients with Brucella melitensis bacteremia

The sensitivity of the standard agglutination test (SAT) for detecting brucellosis was determined in 264 Israeli patients from whom a positive blood culture for Brucella melitensis and serology were obtained within ± 1 week. A SAT titer ≥1:160 had a diagnostic sensitivity of 91.7%, whereas raising the cutoff to ≥1:320, as recommended to decrease false-positive rates in endemic areas, reduced the sensitivity to 82.6%. Physicians working in regions endemic for brucellosis should be aware of the limitations of the SAT for detecting patients with the disease.