虎红平板凝集试验、标准试管凝集试验、酶联免疫吸附方法在布鲁氏菌病中检测的特性及诊断效能

目的 研究虎红平板凝集试验（Rose-Bengal plate agglutination test, RBPT）、标准试管凝集试验（standard tube agglutination test, SAT）、酶联免疫吸附（enzyme-linked immunosorbent assay, ELISA）方法在布鲁氏菌病中检测的特性及诊断效能。方法 选取兴安盟人民医院2020年3月—2021年5月接诊的且有完整记录的489例疑似布鲁氏菌病患者。以接触史+临床症状+血清学检测/布鲁氏菌分离培养为诊断金标准,分析SAT、RBPT和ELISA对布鲁氏菌病的诊断价值。结果 489例疑似患者确诊阳性183例（37.42%）,而RBPT、ELISA、SAT分别检出阳性234例（47.85%）、148例（30.27%）、195例（39.88%）;RBPT诊断布鲁氏菌病的灵敏度、特异度、阳性预测值、阴性预测值、准确度分别为95.08%、80.39%、74.36%、96.47%、85.89%,ELISA诊断布鲁氏菌病的灵敏度、特异度、阳性预测值、阴性预测值、准确度分别为78.69%、98.69%、97...     

随州市羊种3型布鲁氏菌首发病例的病原学检测

目的分离和鉴定布鲁氏菌,为病例的诊断、治疗及疫情的控制提供科学依据。方法虎红平板凝集试验（RBPT）、试管凝集试验（SAT）、酶联免疫吸附试验（ELISA）检测病人血清中特异性抗体,用血清凝集试验、噬菌体试验、PCR试验法对菌株进行分型鉴定。结果虎红平板凝集试验出现絮状抗原抗体结合物100%凝集为阳性;试管凝集试验结果为阳性1：200（＋＋＋）;发病期血清IgM抗体阳性、IgG阴性;M5布鲁氏菌疫苗株和病人菌株提取的基因组进行扩增和电泳,均在223和731bp附近出现阳性条带,而711、976和285 bp处无条带,说明该病人分离出的菌株为布鲁氏菌属羊种布鲁氏菌;经CO2、硫化氢、染料抑菌、菌株表面抗原凝集、噬菌体裂解试验,鉴定该菌株为羊种3型。结论随州市布病首发病例病原体为羊种3型布鲁氏菌,应加强布病监测,采用RBPT、SAT、ELISA、PCR等方法可快速检测病原体,为临床诊断、疫情控制提供依据。     

胶体金法在内蒙古布鲁氏菌病病区的试验

目的评价胶体金法在内蒙古布鲁氏菌病监测地区的现场应用效果。方法选择内蒙古布鲁氏菌病监测地区重点人群,采用虎红平板凝集试验（RBPT）、试管凝集试验（SAT）和胶体金法进行检测,并对血清学结果进行对比分析。结果共检测1088份疑似布病患者血清,在RBPT、SAT检测均为阳性的645例中胶体金法检测为阳性的483例、阴性162例,吻合率为74。8％;在RBPT、SAT检测均为阴性或达不到布病诊断标准的443例中胶体金法检测为阳性的11例,假阳性率为2．5％。结论布鲁氏菌抗体检测试剂（胶体金法）检测布鲁氏菌病与RBPT和SAT检出的阳性布病患者的吻合率不高,该试剂的推广使用价值还需更多实验数据进一步支持。     

The bovine immune response to Brucella abortus IV. Studies with a double immunodiffusion test for antibody against A2

A double immunodiffusion test for precipitins against Brucella antigen A2 was developed and applied to a variety of samples. The A2 precipitins were produced by a heifer infected with B. abortus strain 2308, cattle vaccinated with killed B. melitensis strain H38 or live B. abortus strain 19 and by a dog infected with B. canis. Precipitins were also detected in the second International Standard for anti-Brucella abortus serum, in several anti-B. canis sera and at low levels in one anti-B. ovis serum tested. Antisera produced in calves against Yersinia enterocolitica serotype 0:9 had no anti-A2 activity despite titers greater than or equal to 1/1024 and greater than or equal to 1/80 in standard Brucella agglutination and CF tests, respectively. The test for A2 precipitins lacked specificity as weak reactions were obtained with five of 295 sera from brucellosis-free herds. This test was relatively insensitive, detecting precipitins in only 16 of 24 sera from infected cattle and 27 of 54 sera positive by complement fixation and enzyme labelled antiglobulin tests performed with whole cell and smooth lipopolysaccharide antigens, respectively. The A2 precipitins were detected in nine sera from five cattle, in two infected herds, which were negative by agglutination and complement fixation tests.     

The bovine immune response to Brucella abortus. II. Elimination of some sporadic serological reactions by chelation of divalent cations.

The standard agglutination tests for detecting antibody to Brucella abortus were modified by addition of chelating agents (EDTA and EGTA) to the antigens. Approximately 80% of "singleton" agglutination test reactions, negative on the diagnostic complement fixation test, obtained with cattle sera were eliminated while no decrease in titer was apparent when sera from B. abortus infected or vaccinated cattle were tested.     

珠海市职业人群布鲁氏菌病血清学监测结果

目的 了解珠海市职业人群布鲁氏菌病感染情况.方法 采用血清流行病学监测方法,选取珠海西部地区与牲畜及畜产品有接触的重点人群,设计问卷调查表进行流行病学调查,并采集血清进行虎红平板凝集试验.采用单因素分析和多因素Logistic回归分析对职业危险因素进行探讨.结果 共监测226个职业人群,采集到226份标本,检出阳性标本23份,阳性率为10.2%.多因素Logistic回归模型结果显示,幼畜接生员OR=6.00(1.11～32.46)是危险因素,使用消毒液洗手OR=0.32(0.12～0.90)是保护因素,差异有统计学意义.结论 珠海市畜牧业从业人群中,幼畜接生员工种感染布鲁氏菌病的危险性大,使用消毒液洗手具有保护作用.     

2008年四川省阿坝州布鲁杆菌病调查结果分析

目的了解四川省阿坝州布鲁杆菌病(简称布病)感染状况,分析感染来源和传播途径。方法按全国布病监测方案要求,对重点人群进行流行病学调查和血清学检查(试管凝集试验SAT、平板凝集试验PAT);对牦牛、绵羊血清用SAT试验、虎红平板凝集试验(RBPT)进行检测。结果2008年阿坝州发现人间布病9例,男性2人,女性7人;流行病学调查430人,人血清530份,阳性13人,检出率为2.45%(13/530)。共检疫牛羊5801头,阳性182头,检出率3.13%(182/5801);其中牛5.00%(146/2922)、羊1.25%(36/2879)。结论阿坝州人畜间布病疫情呈上升趋势,主要原因是牛羊进行检疫、免疫力度不够,病畜作为传染源长期存在,牧民群众缺乏布病自我防护意识。     

Evaluation of different serological techniques in laboratory diagnosis of brucellosis

Diagnosis of brucellosis is vital for early institution of proper therapy as untreated cases may progress to chronic stage. Though the demonstration of the causative agent in blood is considered as the most conclusive test in the diagnosis of brucellosis, isolation of brucella organism by blood culture is relatively low. Hence a number of sensitive and rapid serological tests have been introduced for the diagnosis of brucellosis. In the present study, an attempt was made to compare the efficacies of existing serological tests such as agglutination reaction with newer rapid tests which help in the detection of either specific antigen or antibody. The study included specimens from 80 patients clinically suspected to be suffering from brucellosis and 20 apparently healthy controls. All serum samples were subjected for evidence of brucellosis by five serological tests viz, standard tube agglutination test, 2-mercaptoethanol test, modified antiglobulin test, counter immuno-electrophoresis and passive haemagglutination test for antibody detection and two serological tests viz, counter immunoelectrophoresis and latex agglutination test for antigen detection. Eighty blood samples were processed for microbiological evidence of brucellosis and yielded only 8 isolates of Brucella melitensis of biotype 1. By standard tube agglutination test, 25 sera showed titre of brucella agglutinins equal to more than the diagnostic titre (i.e., more than or equal to 160 IU). Counter immuno-electrophoresis test and latex agglutination showed presence of antigen in 3 and 4 blood culture negative cases respectively.     

Haemophilus somnus: a comparison among three serological tests and a serological survey in beef and dairy cattle

Serological tests for the detection of antibodies against Haemophilus somnus were carried out in herds of beef and dairy cattle using three different techniques: agglutination, complement fixation and counterimmunoelectrophoresis. The agglutination test appeared to detect more seroreactors than the complement fixation and counterimmunoelectrophoresis tests. Results of the three tests indicated that there were more positive reactors in beef cattle and dairy cattle from infected herds than in dairy cattle from clinically normal herds.     

Sensitivity and specificity of an enzyme-linked immunosorbent assay for the detection of infectious bovine rhinotracheitis viral antibody in cattle

An enzyme-linked immunosorbent assay was developed to detect bovine serum antibody to infectious bovine rhinotracheitis virus. The specificity of this assay in 304 bovine sera, collected from an infectious bovine rhinotracheitis virus-free herd, was 100%; in sera from 62 cattle inoculated with an intranasal vaccine, its diagnostic sensitivity was 27.4% at one month and 100% at six months, postvaccination. In 303 bovine sera with standard serum neutralizing antibody titers of greater than or equal to 1:2 it showed 100% sensitivity; and in 463 random diagnostic samples, comparative tests indicated that enzyme-linked immunosorbent assay detected more seropositive animals (61.6%) than the standard serum neutralizing test (49.9%). The enzyme-linked immunosorbent assay method was considered to be technically superior as a routine diagnostic test for the detection of infectious bovine rhinotracheitis viral antibody in bovine sera.     

南宁市职业人群布鲁菌感染状况调查

目的了解南宁市职业人群布鲁菌感染状况,为开展科学防治工作提供依据。方法采用试管凝集试验对南宁市南、北2大奶场的饲养员、挤奶工和农贸市场的生畜肉的销售员进行3年的布鲁菌抗体血清学跟踪检测。结果636人份血清中检出阳性37份,阳性率为5．81％,南边奶场和农贸市场畜肉类销售员均为阴性。阳性感染者集中在北边奶场。且数量呈逐年递增趋势。检测和回访中发现阳性感染者血清抗体滴度以1：100～200者占97．30％（36／37）,达到1：400仅1份,感染者未见有临床症状或有不适。结论本市职业人群布鲁菌感染者的抗体阳性滴度不高,几乎均无临床症状。控制家畜布鲁菌病的流行是降低人群布鲁菌感染率的关键,同时应继续关注并跟踪隐性感染者的健康状况。     

Antibody development in pigs inoculated with live or killed cultures of Brucella abortus.

As shown by density gradient ultracentrifugation and column chromatography, pigs formed IgM antibodies during the first week following vaccination with Brucella abortus, strain 19. At this time their sera reacted in both plate and tube agglutination but not in complement-fixation tests. A few days later, when IgG antibodies had developed, agglutination titers were still high and some activity was recorded in hemolytic complement-fixation tests. A similar sequence was observed in pigs repeatedly inoculated with phenol-killed suspensions of B. abortus. As the proportion of IgM to IgG antibodies decreased, agglutinin titers fell in relation to complement-fixing titers. In some animals the conglutinating complement absorption test became positive earlier than the plate agglutination.     

深圳市部分重点人群布鲁氏菌病监测情况分析

目的了解畜牧业重点人群布鲁氏菌病(Brucellosis)感染情况和其危险因素,探索宝安区布鲁氏菌病防治对策。方法对2005年2092名职业重点人群(养殖业、乳业、屠宰)中532人采用血清作试管凝集试验进行检查。结果职业重点人群感染率0.38%(2例)。结论畜牧业重点人群存在布鲁氏菌病感染的概率,应视为宝安区人间布病感染不可忽视的传染源,把从事畜牧业职业人群作为宝安区今后人间布鲁氏菌病监测的重点。     

A comparison of some serological tests for bluetongue virus infection.

The plaque neutralization, complement fixation, and agar gel precipitin tests were compared by measuring bluetongue virus antibody in 137 serums from experimental animals (cattle and sheep) and suspected field reactors (cattle and deer). In general, the tests agreed well with each other. Plaque neutralization titers began earlier than the other two and went much higher than the complement fixation titers. Plaque neutralization titers usually peaked between two and three weeks after exposure and complement fixation titers from four to six weeks. The greater sensitivity of the plaque neutralization test allowed the detection of all complement fixation and agar gel precipitin reactors whereas occasionally the latter two tests failed to detect plaque neutralization reactors.     

幽门螺杆菌粪便抗原免疫卡在诊断幽门螺杆菌现症感染和判断其在根除治疗中的价值

目的　评价幽门螺杆菌粪便抗原 (HpSA)快速免疫检测卡诊断Hp感染和判断Hp根除疗效的敏感度及特异度。方法　对 80例有上胃肠道症状患者的粪便标本作HpSA快速免疫卡检测 ,80例中 4 0例为接受胃镜检查而未接受过Hp根除治疗者 (A组 ) ,A组中又有 2 6例同时采用HpSA免疫卡作胃黏膜组织抗原检测 ;另外 4 0例为Hp根除治疗后的复查患者 (B组 )。诊断Hp感染和判断Hp根除的“金标准”采用Hp培养、快速尿素酶试验和Warthin Starry银染或者13 C 尿素呼气试验(UBT)进行检测 ,“金标准”是指上述 3项中 2项阳性或者Hp培养阳性 ,全部检查阴性认为Hp阴性 ,未作胃镜者以13 C UBT检查为标准。结果　 80例患者中按“金标准”诊断Hp阳性 36例 ,阴性 4 4例 ;HpSA检测阳性 39例 ,阴性 4 1例 ,假阳性 3例 ,与“金标准”比较 ,其敏感度和特异度分别为 :10 0 %、93 2 % ;若A组与B组分别比较 ,A组分别为 10 0 %和 86 7% ;B组分别为 10 0 %和 96 6 % ;HpSA检测组织抗原的敏感度和特异度分别为 92 3%和 92 3%。结论　HpSA快速免疫检测卡检测粪便Hp抗原不仅可以用于临床Hp现症感染的诊断 ,而且也可用于Hp根除治疗的结果判断     

Evaluation of the complement fixation test for the diagnosis of pleuropneumonia of swine caused by Haemophilus pleuropneumoniae

Evaluation of diagnostic sensitivity and specificity was based on test results of 346 sera from pigs known to be infected and 139 sera from pigs known not to be infected. All sera were tested with a monospecific antigen (serotype 1) and a polyspecific antigen (serotypes 1-5). The sensitivity of the polyspecific antigen was approximately 85% at serum dilution 1:2 and was significantly higher than the monospecific antigen at all serum dilution levels. The specificity of the two antigen preparations was not significantly different at any dilution and increased from approximately 78% to 1:2 to 100% at 1:128. When pigs from herds with unknown incidence of infection were studied, it was found that a high proportion seroconverted, presumably as a response to subclinical infection. However, the antibody titres waned rapidly. This indicated that seroreaction expresses current or recent infection. Thus, the complement fixation test provides a reliable means of diagnosing pleuropneumonia of pigs and might be useful as a tool to control this disease.     

应用ROC曲线评价耐甲氧西林葡萄球菌的检测方法

目的：通过对耐甲氧西林葡萄球菌（MRS）的常用检测方法及实验条件进行评价，选择特异性和敏感性较好的方法和理想的实验条件。方法：以PCR法检测葡萄球菌的mecA基因为“金标准”，比较PBP2a胶乳凝集试验法、头孢西丁（FOX）琼脂筛选法、苯唑西林（OXA）琼脂稀释法（MIC）、FOX纸片扩散法、OXA琼脂筛选法、OXA纸片扩散法检测MRS的敏感性和特异性。同时应用ROC曲线对实验方法及不同的实验条件进行评价。结果：PCR法共检出79株mecA基因阳性菌株，阳性率为73．8％。以PCR法为“金标准”，上述6种方法的ROC曲线下面积分别为0．994，0．964，0．955，0．946，0．932，0．862。结论：选择最佳实验条件，用两种表型检测方法作平行检测，对可疑结果用PBP2a胶乳凝集试验或PCR法确诊。     

海南2例少见布鲁菌感染患者的诊疗

本文报道了检验医师参与海南地区2例布鲁菌感染患者的诊断、治疗和疗效评价。两例患者均出现畏寒、发热现象,其中一例伴有四肢乏力、食欲不振,另一例伴有明显头痛。血培养革兰染色镜检见革兰阴性短小杆菌,呈“细沙样”,柯氏染色待检菌为阳性(红色),大肠埃希菌为阴性(蓝色),15 min尿素酶强阳性,具备布鲁菌感染的特点。血清试管凝集试验(SAT)效价分别为1:200及1:600。考虑到患者曾有牛羊接触史,结合发病特征及实验室检查,确诊为布鲁菌病。经利福平(600 mg/d)联合多西环素(200 mg/d)正规治疗6周后患者恢复良好。     

A study of complement-fixation and serum agglutination tests on vibrio fetus infected and immunized cattle

Complement-fixation (CF) and tube agglutination (TA) tests for demonstration of Vibrio fetus antibodies were conducted on the sera of three groups of ten heifers. One group was vaccinated subcutaneously with a commercial V. fetus var venerealis bacterin and challenged intra-utero, at the external os cervicus one month later; the second was infected only and the third vaccinated only.The vaccinated cattle developed high CF serum titers, but no such increase was observed in animals infected only. A moderate increase in serum antibody titers was demonstrated by the TA test following either infection or vaccination; although titers observed were not higher than those observed in the sera of some apparently normal uninfected animals. The group receiving both vaccine and challenge was the only one in which significant serum antibody titers were demonstrable by the TA test. The sera of these animals also had significant titers in the CF tests.The CF and TA tests detected serum antibodies produced by the parenteral inoculation of V. fetus antigen. These two tests were of limited value in detecting serum antibodies from animals with genital V. fetus var venerealis infection, although the formation of local antibodies was demonstrable by the vaginal mucus agglutination test.     

SPA—乳凝抑制试验检测抗HBC抗体

作者用SPA—聚苯乙烯胶乳作为载体,研制抗HBc乳凝试剂,通过乳凝抑制试验(LAIT)检测抗HBc抗体。结果显示,LAIT具有良好的敏感性(检出5mg/L抗HBc)和特异性(90.3％)。试剂稳定、方法简便、快速。与酶联免疫吸附试验(ELISA)比较,在HBsAg阳性的44份血清中,抗HBc的检出率:LAIT为50％,ELISA为54.5％两者相似,总符合率是86.4％。因此,作者认为LAIT是检测抗HBc的好方法。