牛乳布氏杆菌抗体ELISA检测试剂盒与血清凝集试验检测方法的比较

布氏杆菌病（Brucellosis）是由布氏杆菌引起的一种重要的人畜共患传染病。该病广泛分布于世界各地，目前在我国人、畜间仍有发生，给畜牧业生产和人类健康带来了严重的危害。现在我国对奶牛布氏杆菌病的检疫常用虎红平板凝集试验（RBPT）、试管凝集试验（SAT）和补体结合试验（CFT）等血清学方法，但这些血清学方法有的特异性和灵敏性不高，有的操作繁琐。     

2016—2018年酒泉市肃州区羊布氏杆菌病血清学监测

研究旨在对甘肃省酒泉市肃州区羊布氏杆菌病流行情况进行血清学调查,为该病的防控和净化提供参考。在2016年3月—2018年12月对肃州区不同地区羊场采集羊血清样品共3203份,应用虎红平板凝集试验对血清样品进行布氏杆菌病的血清学检测。结果显示:在被调查的3203份血清样品中,7份检测为布氏杆菌病阳性,阳性率0.22%,其中2016—2018年羊布氏杆菌病阳性率分别为0.48%、0.29%和0.0%;不同季节的检出率差异不明显,春季、夏季、秋季和冬季羊布氏杆菌病检出率分别为0.32%、0.19%、0.10%和0.35%,提示布氏杆菌病在肃州区羊群存在流行情况,应该加强对该病的监测工作。     

抗凝剂对布氏杆菌病检测结果的影响

为了解血浆、血清对布氏杆菌病虎红平板凝集试验检测结果是否有影响，试验取20只成年羊的颈静脉血，分别进行了虎红平板凝集试验和试管凝集试验，结果发现：血清样品布氏杆菌病RBPT、SAT的检测结果均为阴性；血浆样品布氏杆菌病RBPT的检测结果均为阳性，SAT的检测结果均为阴性。     

奶牛布氏杆菌病的血清学诊断

为了解虎红平板凝集试验(RBPT)、试管凝集试验(SAT)和酶联免疫吸附试验(ELISA)三种血清学方法在奶牛布氏杆菌病诊断方面的实际应用情况,本试验采集了10份疑似奶牛布氏杆菌病的血清样本,同时采用上述三种血清学诊断方法进行了对比研究。结果显示,RBPT、SAT和ELISA的阳性检出率分别为70%、80%和90%,以ELISA的阳性检出率为最高。因此,具备特异、灵敏、快速等优点的ELISA血清学诊断方法更适合奶牛布氏杆菌病的抗体检测及其研究。     

细胞壁抗原用于布氏杆菌病诊断试验的研究

用培养的布氏杆菌菌体，通过超声波裂解、反复离心制备出布氏杆菌细胞壁抗原。将细胞壁抗原作1：128稀释，用作牛种布氏杆菌酶联免疫吸附试验（ELISA)的抗原；将布氏杆菌细胞壁抗原作1：32稀释，用作平板凝集试验抗原；把细胞壁抗原作1：16稀释用作试管凝集试验抗原，分别建立了牛布氏杆菌ELISA试验、平板凝集试验和试管凝集试验。用这3种方法，检测已知200份平板凝集试验阴性血清，5份平板凝集试验阳性血清。结果5份阳性血清在平板凝集试验、试管凝集试验和ELISA试验中均为阳性；200份阴性血清在平板凝集试验和试管凝集试验中均为阴性，在ELISA试验中有1份为阳性?试验证明，细胞壁抗原，既能用于传统的平板凝集试验和试管凝集试验，又能用于ELISA试验。     

全乳环状试验检测奶牛布氏杆菌病的试验探讨

用全乳环状试验 (ABRT)、试管凝集试验 (SAT)、虎红平板凝集试验 (RBPT) 3种血清学方法对南京地区 49头奶牛的血清和乳汁进行了布氏杆菌病的检测试验 ,并用ABRT法检测 2 0 0 0多份乳样。试验结果证明 ,ABRT法具有较高的敏感性和较强的特异性。该法操作简便、易行、快速、结果可靠、准确性高 ,可以在布氏杆菌病实际检疫中进行推广应用 ,并可替代SAT法和RBPT法     

循化县山羊布氏杆菌病衣原体病的血清学调查

为了了解循化县山羊布氏杆菌病（以下简称布病）和衣原体病的感染情况，笔者于１９９６年布氏杆菌试管凝集试验（ＳＡＴ）和衣原体间接血凝试验（ＩＨＡ）对６８６只１岁以上的山羊进行了血清学调查，现将结果报告如下。１　　材料与方法１．１　　被检血清：采自循化县未免疫的一岁以上的山羊。１．２　　诊断用试剂：布氏杆菌ＳＡＴ抗原，阳性血清和阴性血清系成都生物制品厂提供，批号为９６０４１８，稀释液用市售０．９％生理盐水。衣原体ＩＨＡ抗原阳性血清、阴性血清和特定稀释液均系中国农业科学院兰州兽医所提供，批号为９６０５２…     

利凡诺尔血清凝集试验诊断猪布氏杆菌病的报告

利凡诺尔血清凝集试验诊断猪布氏杆菌病的报告梁杏娴，吴福材（广东省兽医防疫检疫站）（中国兽药监察所）尹锦霞，何赐贤（广州交通兽医检疫站）（清远市畜牧局）布鲁氏菌病的血清学诊断方法很多，补体结合试验波认为是最可靠的诊断方法。但猪血清有抗补体作用，所以用补...     

乳畜和高危人群布氏杆菌病血清学监测及综合防治措施

为了解乳畜与高危职业人群布氏杆菌病感染现状,探索人间和畜间布氏杆菌病综合防治措施,应用国家标准试管凝集法,对云南省某县126份奶牛、奶山羊饲养人员、村人医、兽医和屠户等高危职业人员血清样品进行检测,对85份混合牛奶、羊奶样品进行ELISA方法检测.结果表明：人血清布氏杆菌病血清学阳性率3.97％,混合奶样品布氏杆菌病血清学阳性率7.06％,通过布氏杆菌病发生的相关因素分析,提出布氏杆菌病综合防治措施.     

吉林、黑龙江两省梅花鹿布氏杆菌病的血清学调查

应用血清学方法,对我国吉林、黑龙江两省梅花鹿成年鹿群未注射布氏杆菌病疫苗的874份血清进行了血清抗体检测,其抗体阳性率分别为8.87%、2 6.39%。这一数据表明,吉林省和黑龙江省的鹿群中存在着布氏杆菌病的感染。同时只对不同性别的梅花鹿进行了血清抗体检测,结果表明,梅花鹿公鹿和母鹿对布氏杆菌均易感。此研究为今后有效的预防和治疗布氏杆菌病提供了临床理论依据     

布鲁氏菌4种高通量抗体检测方法的比较研究

旨在科学选择和使用布鲁氏菌抗体检测方法,推动布病诊断试剂标准化。本研究用布病阳性血清标准品测定了国家/OIE布鲁氏菌病参考实验室开发的布鲁氏菌荧光偏振（FPA）抗体检测试剂盒、动物布鲁氏菌病竞争ELSIA （cELISA）抗体检测试剂盒、牛布鲁氏菌病间接ELISA （iELISA）抗体检测试剂盒和改进的微量补体结合试验（mCFT）等4种方法的灵敏度。通过对已知阴、阳性血清样品的检测,比较了各检测方法的敏感性和特异性,并用临床样本进一步比较了各种方法检测结果的吻合性。结果表明,4种方法检测的灵敏度基本一致,当布病阳性血清标准品按1∶20稀释（即50 IU·mL^-1）时均检测为阳性,1∶40稀释（即25 IU·mL^-1）时均检测为阴性。FPA、cELISA、iELISA和mCFT方法的敏感性分别为97.14%、100.00%、100.00%、98.57%,特异性分别为96.34%、95.12%、97.56%、100.00%。对315份临床样本的检测结果显示,各方法之间的符合率均高于90.00%,其中iELISA、FPA、cELISA与mCFT符合率分别为97.14%、96.83%、92.70%;FPA、cELISA与iELISA符合率分别为95.24%、93.65%;FPA与cELISA符合率为91.43%。iELISA、FPA、mCFT 3种方法之间吻合性最高,cELISA与其他3种方法之间的吻合性略低。     

Validation and standardization of virus neutralizing test using indirect immunoperoxidase technique for the quantification of antibodies to rabies virus

A virus neutralizing test using an indirect immunoperoxidase technique (VNT-IIP) for rabies has been developed for the titration of dog and cat serum samples in Japan. The VNT-IIP has the advantage that results obtained can be viewed by the naked eye. The purpose of this study was to validate the VNT-IIP and compare it with one of the international standard methods, the fluorescent antibody virus neutralization test (FAVNT). The VNT-IIP showed satisfactory repeatability, high analytical specificity and good accuracy. Regarding the comparison between the VNT-IIP and the FAVNT, the VNT-IIP showed good agreement (91.9%), high sensitivity (92.8%) as well as specificity (87.0%) and good correlation (r = 0.92). As described above, the validation of the VNT-IIP was satisfactory and the performances of the test proved to be equivalent to those of an international standard method.     

几种布鲁氏菌病血清学诊断方法的比较研究

疑似布病感染的牛场采集牛奶和全血进行细菌分离,采集血清用虎红平板凝集试验(RBT)、试管凝集试验(SAT)、iELISA、cELISA进行抗体检测;采集免疫牛场和部分免疫羊场血清430份进行国内4个厂家生产的RBT抗原比对实验,并且选择特异性最高厂家的RBT抗原与SAT、iELISA和cELISA同时进行抗体检测。结果表明4个厂家生产的RBT抗原检测结果的一致性较差,RBT和SAT与加拿大布病参考实验室提供的iELISA和cELISA试剂盒检测结果相比一致率较高,但前两者均有较高的假阳性和假阴性。通过对比试验得出:在布病检疫时可选择特异性好的RBT抗原进行初筛,阳性结果用iELISA或cELISA进行确诊。     

不同方法检测鸡白痢沙门氏菌抗体消长规律的比较研究

为比较不同方法检测鸡白痢沙门氏菌抗体消长的规律，以便为种鸡场净化提供科学指导，本研究以鸡白痢沙门氏菌活菌和灭活免疫原分别接种SPF鸡，采用平板凝集、微量凝集和ELISA试验定期检测血清中的特异性抗体，并以Kappa检验判定不同检测方法之间的一致性程度。结果显示，平板凝集试验在接种后检出抗体阳转的时间早于ELISA，但ELISA检出抗体阳性的持续时间更长，且更符合抗体消长规律；3种检测方法的结果之间仅具有微弱一致性（Kappa系数为0.002~0.295）。本研究结果表明，不同鸡白痢沙门氏菌抗体检测方法之间存在较大差异，但从总体来看，ELISA无假阳性干扰，检出抗体的持续时间较长，更符合抗体消长规律，在单一鸡白痢沙门氏菌感染的情况下，更有利于种鸡场对该病进行净化。     

乳胶凝集试验与血清中和试验检测猪伪狂犬病血清抗体的 …

应用血清中和试验（ＳＮＴ）和伪狂犬病乳胶凝集试验（ＬＡＴ）诊断试剂盒对两种伪狂犬病是性血清、伪狂犬病病毒（ＰＲＶ）高兔血甭及６０份被检猪血清进行了ＰＲＶ抗体效价测定和相关性分析,两种方法测得的抗体效价之间呈强相关性（ｒ＝０．９６）,且ＬＡＴ效价比ＳＮＴ一般高出一个滴度;能干为自３５个猪场的４１４份猪血清进行了ＰＲＶ抗体检测,并与ＳＮＴ检测结果进行了对比,结果在ＳＮＴ检测为阳笥的１７１份血清中,ＬＡ     

Comparison of results from five serologic methods used for detecting Brucella abortus antibody activity in coyote sera

A total of 423 serum samples representing 94 coyotes which were wild trapped in east Texas were used to compare the serologic results from five different methods for detecting antibodies to Brucella abortus. The sera were tested for Brucella spp. antibody activity by the Card (CARD), rivanol precipitation (RIV), standard agglutination tube (SAT), cold complement fixation test (CF), and enzyme linked immunosorbent assay (ELISA) methods. Each serum sample selected for this comparison demonstrated antibody activity by one or more of the five serologic methods. When the serologic results of the five different methods were compared, 143 sera were positive according to the CF test and agreement was 67.1-70.6% with CARD, RIV and SAT. The maximum agreement for CF positive was with CARD (70.6%) and the lowest agreement fro CF negative was also with CARD (56.4%). Agreement among the serologic methods for the SAT positive ranged from 69.1% (CARD) to 72.7% (RIV). Agreement between SAT and ELISA was poor with only 38.1% agreement for SAT positive and 11.3% agreement for SAT negative. Agreement between methods for CARD positive sera was poor, with a low of 43% for both SAT and ELISA, and a high of 55.6% for RIV. Agreement between methods for 149 RIV positive sera was 83.2% for CARD, 67.8% for SAT, 64.4% for CF and only 50.3% for ELISA. Agreement between methods for ELISA positive results ranged from 49.0% for RIV to 62.7% for CARD.(ABSTRACT TRUNCATED AT 250 WORDS)     

检测猪细小病毒血清抗体乳胶凝集试验方法的建立及初步应用

将猪细小病毒在IBRS-2细胞上同步培养增殖,待出现细胞病变后,反复冻融,收获病毒液,用甲醛灭活。随后用经硫酸胺沉淀、透析后浓缩的细小病毒液进行方阵滴定,选择致敏乳胶的最佳条件,制成细小病毒乳胶凝集试验（LAT）抗原。抗原与细小病毒阳性血清反应出现肉眼可见的凝集颗粒,而与生理盐水、PBS、犊牛血清、猪瘟、衣原体、口蹄疫、伪狂犬病、弓形体及萎缩性鼻炎等阳性血清不出现凝集现象。用所建立的细小病毒乳胶凝集试验（LAT）与血凝抑制试验检测了203份猪血清,其中血凝抑制抗体阳性为161份,阴性为42份,阳性率为79.31％;乳胶凝集抗体阳性为150份,阴性为53份,阳性率为73.89％,两种方法阳性符合率为93.16％,经统计学检验P＞0.05,两者差异不显著。结果表明,乳胶凝集试验可以作为临床上大量血样进行血凝抑制试验前的初筛,具有简便、准确的优点,在检测细小病毒（PPV）抗体上具有较好的应用前景。     

Evaluation of different enzyme-linked immunosorbent assays for the diagnosis of brucellosis due to Brucella melitensis in sheep

Six serological assays for the diagnosis of ovine brucellosis, due to Brucella melitensis were evaluated. Reference serum samples from sheep of known B. melitensis infection status (n = 118) were assessed using the Rose Bengal test (RBT), complement fixation test (CFT) and four commercial enzyme-linked immunosorbent assays (ELISAs), including two indirect ELISAs (iELISAs), one competitive ELISA (cELISA) and one blocking ELISA (bELISA).The highest differential positive rates (DPR) were obtained with the cELISA and bELISA, while the lowest DPR was estimated using iELISAs. A latent class analysis was performed to estimate the accuracy of the CFT, RBT and bELISA using 1827 sera from sheep undergoing testing as part of a surveillance and control programme. Lower sensitivity and specificity were obtained for the three serological tests when the field samples were used. A higher DPR was achieved by the CFT, compared to bELISA and RBT. The results suggest that ELISAs, and particularly the bELISA, might be suitable for inclusion in the European Union legislation on intra-community trade for diagnosing B. melitensis infection in sheep, as it has a similar test performance compared to the RBT.     

牛结核病不同检测方法的比较研究

[目的]为达到迅速准确诊断牛结核病的目的,[方法]用2年时间对我国最常用的牛结核病检测方法进行了对比研究.[结果]试验结果表明,国标皮内变态反应试验(PPD)与皮内变态反应比较试验(SICTT)有较高符合率;国标皮内变态反应试验(PPD)阳性敏感性高于皮内变态反应比较试验(SICTT);皮内变态反应比较试验(SICTT...     

Comparison of the Q-fever complement fixation test and two commercial enzyme-linked immunosorbent assays for the detection of serum antibodies against Coxiella burnetii (Q-fever) in ruminants: Recommendations for use of serological tests on imported animals in New Zealand

Abstract

AIM: To make valid recommendations on the use of serological test methods for the detection of serum antibodies in ruminants against Coxiella burnetii (Q-fever), by comparing the performance of the complement fixation test (CFT) and two ELISA, and by identifying reasons for discrepancies between the test methods.

METHODS: A total of 73 serum samples from infected cattle, 69 from infected goats, and 100 samples from non-infected cattle and 57 samples from non-infected sheep, as well as 95 samples from infected cattle herds (mix of seropositive and seronegative samples), were tested using the CFT, the IDEXX ELISA (I-ELISA) and the Pourquier ELISA (P-ELISA). A mixed panel of 12 serum samples from sheep from inter-laboratory proficiency testing (proficiency panel) was also tested using the CFT and both ELISA, and further investigated using IgG- and IgM-specific ELISA.

RESULTS: Generally, the two commercial ELISA were more sensitive than the CFT for the detection of infected ruminants. Good agreement between ELISA for positive and negative results was found for samples from the infected herd, while results for the positive panels varied between the two ELISA. For the total of the positive serum panels, the I-ELISA detected 95% of samples as positive or suspicious, while the P-ELISA detected only 81%. In the P-ELISA, more samples were considered suspicious (18%) than in the I-ELISA (14%). All sera from noninfected sheep and cattle tested negative in the serological test methods employed, except for one positive sample from a sheep in the P-ELISA. Further investigation revealed that a CFT-positive but ELISA-negative result was due to high IgM and low IgG reactivity.

CONCLUSIONS: The two commercial ELISA were more sensitive than the CFT in all panels from infected ruminants. However, they could only detect IgG. The I-ELISA should be the serological test method of choice for cattle, sheep and goats for import testing of animals into New Zealand because it was more sensitive than the P-ELISA and was equally specific to the PELISA and the CFT. For other animal species, such as deer and camelids, the CFT should still be used since none of the ELISA has been evaluated for these species. This study has shown that the two commercial ELISA will detect the majority of infected ruminants but may miss animals that have not developed an IgG response.