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乳胶凝集试验检测鸡新城疫病毒血清抗体的研究 总被引：13，自引：1，他引：12
用硫酸铵沉淀、透析浓缩的鸡新城疫病毒（ＮＤＶ）抗原致敏乳胶,然后所鸡新城疫阳性血清进行方阵滴定,选择出最佳致敏条件并制成新城疫病毒乳胶抗原,由此建立超乳胶凝集试验（ＬＡＴ）,用来检测新城疫血清抗体;用所建立的乳胶凝集试验（ＬＡＴ）和血凝抑制试验（ＨＩ）同步检测６９份鸡血清,结果乳胶凝集试验阳性６２份,阴性７份;血凝抑制试验阳性６６份,阴性３份,两者的总符合率为９４．２％（６５／６９）,且两者的检出率基本一致。结果表明,乳胶凝集试验具有操作简便、快速、敏感性高、特异性强且可用于现场检测等优点,是一种适合基层单位用来检测鸡新城疫病毒血清抗体的新方法。 相似文献
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重组M蛋白-乳胶凝集试验检测PRRS病毒血清抗体的研究 总被引：19，自引：0，他引：19
利用纯化的 PRRS病毒重组 M蛋白致敏乳胶制成乳胶抗原 ,成功地建立了一种检测 PRRS病毒血清抗体的乳胶凝集试验 (L AT)诊断方法。用制备的乳胶 M抗原分别检测猪瘟、猪伪狂犬病、猪细小病毒病、猪弓形体病、猪衣原体病、猪乙型脑炎阳性血清 ,结果均为阴性 ,无交叉反应 ,说明建立的 L AT方法具有良好的特异性。用建立的乳胶凝集试验方法与国外IDEXX公司 PRRS病毒抗体检测试剂盒同时对 76份猪血清样本进行检测 ,结果表明建立的 L AT方法的特异性和敏感性均为 95 % ,两种方法的总符合率为 87% ,检出率基本一致。研究结果表明 L AT方法具有操作简便、快速、敏感性高、特异性强、价格低廉且可用于现场检测等优点 ,是一种适合基层兽医单位用于 PRRS病毒血清抗体检测的新方法 相似文献
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采用Ni-NTA His.Bind方法纯化pET-28a-ApxⅣ2表达蛋白,以羧化乳胶为载体,经化学交联法致敏乳胶,对致敏条件优化后进行质量检测。结果显示,在pH 4.7的PBS缓冲液中,乳胶浓度为2%、蛋白质量浓度为1.5g/L,偶联8h后制备的致敏乳胶凝集反应最好,致敏乳胶抗原无自凝现象;与副猪嗜血杆菌、猪瘟病毒、猪伪狂犬病病毒、猪繁殖与呼吸综合征病毒、大肠杆菌和多杀性巴氏杆菌阳性血清不发生凝集;批内重复、批间重复试验很稳定;阳性血清敏感性可达1∶32。对比试验中与ELISA方法的符合率为80%,2种方法检测的结果基本相符。结果表明,试验初步建立了检测猪胸膜肺炎放线杆菌ApxⅣ毒素抗体的乳胶凝集方法,为猪传染性胸膜肺炎的快速诊断提供了技术支撑。 相似文献
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以结核分枝杆菌为原料,提取菌体蛋白和多肽。用Lowry法检测菌体蛋白、多肽的含量,并对菌体蛋白、多肽进行Tricine-SDS-PAGE,分析其组分。将所提取的菌体蛋白和多肽分7组作用于鸡体,即菌体蛋白高、中、低剂量组,透析多肽组,未破碎全菌体组,菌体蛋白注射组和对照组,除注射组外,其余6组均采用口服给药。各组试验鸡于给药后7、14、21和28 d 4次屠宰,分别检测新城疫抗体滴度和外周血淋巴细胞转化率。结果表明:不同批次提取的菌体蛋白、多肽含量相对稳定,没有明显差异;组分分析表明,菌体蛋白由多种组分构成,分子量为6.5~66 ku;动物试验结果表明:结核分枝杆菌菌体蛋白试验组均能提高鸡体的细胞免疫和体液免疫功能,尤以菌体蛋白注射组和菌体蛋白中剂量口服组诱导机体免疫效果较好。 相似文献
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用一种新方法测定传染性法氏囊病病毒的特异性抗体，并对抗体效价与保护力的相关性进行了研究，对不同抗体效价的鸡进行攻毒，用乳胶凝集抑制试验测定抗体效价，该法对检测ＩＢＤＶ ＶＰ２特异性抗体快速，简便，当免疫鸡的ＬＩ效价达１：２或更高时，几乎所有的鸡均可抵抗强毒ＩＢＤＶ的攻击，相反，带有母源抗体的鸡，即使其ＬＩ效价达１：４，用同样方法攻击，仍有半数的鸡易感。 相似文献
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番鸭呼肠孤病毒乳胶凝集试验检测方法的建立 总被引：1，自引：0，他引：1
建立了用纯化的抗番鸭呼肠孤病毒(DRV)抗体致敏乳胶检测DRV抗原的方法。通过试验，确定抗体致敏乳胶的最佳浓度为1：10(IgG蛋白浓度为0．4242mg／ml)。最佳致敏温度为37℃。最佳致敏时间为120min。用所建立的方法对人工感染的1日龄雏番鸭30只进行粪便检测。结果表明。感染后第3d粪便中即可检测到病毒．直至人工感染鸭全部死亡都可从粪便中检出病毒抗原；对同样人工感染的30只1日龄雏番鸭的心、肝、脾病料用所建立的方法检测，结果表明．感染后第4d，2／3番鸭脾病料检出阳性；感染后第5d，2只死亡鸭中有1只肝病料检出阳性，脾病料2只都呈阳性；此后的病死鸭脾和肝病料均为阳性，而心则未检出阳性。 相似文献
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用猪链球菌血清2型(Streptococcus suis serotype 2,SS2)的荚膜多糖(Capsular polysaccharide,CPS)致敏经戊二醛处理过的鸡红细胞,优化致敏醛化红细胞的抗原量和时间.结果显示,在37℃条件下CPS(2.2 5 mg/L)致敏鸡红细胞90 min,IHA试验在25℃条件下操作,被检血清IHA效价在1∶8及其以上时判为SS2抗体阳性.应用此方法对猪大肠杆茵、副嗜血杆菌、猪肠球菌、SS1、SS7、SS9、金黄色葡萄球菌、沙门菌、巴氏杆茵阳性血清进行检测,结果表明,SS2抗体均为阴性.对1044份无临床症状的猪血清进行检测,SS2抗体阳性率为61.69％.该方法敏感性高,特异性较强,可用于大规模SS2抗体水平的检测和SS2的血清流行病学调查. 相似文献
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间接酶联免疫吸附试验检测禽流感抗体的最佳工作条件 总被引：26，自引：0，他引：26
禽流感病毒（ＡＩＶ）感染的鸡胚囊液经差速离心后,再经蔗糖密度梯度离心,提纯ＡＩＶ,纯化的ＡＩＶ经ＮＰ４０处理并反复冻融,即为ＡＩＥＬＩＳＡ抗原,用该抗原包被聚苯乙烯微量反应板,将健康鸡ＩｇＧ提纯后免疫兔,制备兔抗鸡ＩｇＧ用过碘酸钠法制备辣根过氧化物酶标记的兔抗鸡ＩｇＧ,确立了间接酶联免疫吸附试验（ＥＬＩＳＡ）检测禽流感抗最适工作条件,即：抗原包被浓度１．９～３．８μｇ／ｍｌ,每孔１００μｌ４℃冰箱 相似文献
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The entire gene encoding microneme protein 3 (MIC3) from Toxoplasma gondii was cloned into the plasmid pGEX-KG and subsequently expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. The recombinant MIC3 (rMIC3) was purified and evaluated in a latex agglutination test (LAT) as the diagnostic antigen for the detection of antibodies to T. gondii in pig sera. The specificity, stability, and reproducibility of the test were examined. No agglutination was found when the sensitized latex beads were mixed with phosphate-buffered saline (PBS), borate-buffered saline (BBS), normal saline, and negative serum samples. There was no cross-reactivity with the standard positive sera of other pathogens. But intense agglutination occurred with T. gondii antibody positive serum samples. In our study, the coincidence rate of tested positive-sera of the LAT with rMIC3-sensitized latex particles and the ELISA with rSAG1 was up to 92.8%, T. gondii specific antibodies were detected by the LAT in all piglets that were experimentally infected with T. gondii tachyzoites from 8 to 42 days after infection. Our results indicated that the rMIC3 based latex agglutination test appears to be suitable for the detection of T. gondii antibodies at the early stage of infection. 相似文献
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Lü J Zhao J Fang L He Q Cao S Chen H 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2006,53(2):59-61
A slide latex agglutination test (LAT) was developed and evaluated to detect serum antibodies against porcine parvovirus. Porcine parvovirus antigen was obtained by 10% PEG-6000 and 0.5 mol/l sodium chloride precipitation, and inactivated by 0.1% methanal. Two per cent suspensions of latex particles (0.5-0.8 microm) were coated by adding an equal volume of porcine parvovirus antigen at 0.34 microg/ml. Repeatability of latex agglutination test was evaluated with a panel of 100 sera using the same and different antigen lots. A good agreement between LAT and haemagglutination inhibit assay was observed. Because of convenience and speed of performance, this method would be used widely in clinic examination. 相似文献
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A rapid semi-quantitative latex agglutination test (LAT) has been standardized for the detection of leptospiral antibodies in serum samples of man and animals. The efficacy of the LAT was compared with the plate enzyme linked immunosorbent assay (ELISA). A total of 276 human serum samples were analyzed by both LAT and ELISA and percentage positives were 84.8 and 85.9%, respectively. Similarly, of 65 animal samples tested, 63.1 and 69.2% positivity were observed in LAT and ELISA, respectively. Even though the ELISA test was slightly more sensitive than LAT, the rapidity, simplicity and economics of the LAT were found to fulfill the requirements of a screening test for leptospiral antibodies. 相似文献
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Relationship of calf antibody status to disease and performance 总被引：1，自引：0，他引：1
Three tests were used to measure the circulating immunoglobulin in 381 purchased calves as they entered a commercial calf-rearing unit. A correlation of 0.64 was found between the zinc sulphate turbidity (ZST) test and a quantitative latex agglutination test (LAT) measuring IgG1 (P less than 0.001). A qualitative version of the LAT related poorly to the quantitative version. The proportion of plasma samples identified by the quantitative LAT as having an IgG1 concentration of less than 5 g/litre which were incorrectly identified as positives (greater than or equal to 5 g/litre) by the qualitative LAT was 0.65. The proportion of plasma samples identified by the quantitative LAT as having a IgG1 concentration of greater than or equal to 5 g/litre which were incorrectly identified as negative (less than 5 g/litre) was 0.11. There was no statistically significant relationship between plasma IgG1 concentration and initial liveweight, subsequent overall daily liveweight gain or disease incidence (P greater than 0.05). Calves treated for infectious disease, particularly respiratory disease after weaning, had statistically significantly lower liveweight gains than healthy calves (P less than 0.001). 相似文献
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A latex agglutination test (LAT) was developed for rapid detection of antibodies to H5 avian influenza viruses (AIVs). The hemagglutinin protein of H5 AIV was covalently linked to carboxylated latex by ethyl-dimethyl-amino-propyl carbodiimide to prepare the sensitized latex beads. The LAT was evaluated with the hemagglutination inhibition (HI) assay as the reference test. The H5-LAT showed a sensitivity of 87.0% and specificity of 88.9% in detecting 126 serum samples from experimentally infected chickens and a sensitivity of 82.5% and specificity of 86% in detecting 587 field chicken serum samples from mostly vaccinated chickens. The agreement ratio between H5-LAT and HI was found to be 87.3% and 83.1% for the two groups of samples, respectively. Difficulty with background agglutination in stored chicken sera was overcome by serum pretreatment with either dried chicken liver powder or dilution buffer containing detergent Tween-20. The H5-LAT has advantages over a previously reported whole-virus LAT in terms of biosafety in preparation, chemical stability, and higher specificity. It is a rapid and simple test suitable for field monitoring of antibodies to H5-type AIV. 相似文献
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A study of pseudorabies virus (PRV)-vaccinated pigs comparing the immune responses detected by the latex agglutination test (LAT) with responses detected by other routine tests for pseudorabies antibodies indicated that LAT was more sensitive than either the enzyme-linked immunosorbent assay (ELISA) or the serum virus neutralization test (SVNT). The LAT detected antibodies sooner than ELISA and SVNT in unvaccinated pigs after challenge with virulent PRV. The specificities of the 3 tests were found to be near 100%. The LAT is a good alternative to SVNT or ELISA for detection of PRV-specific antibodies. 相似文献
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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 总被引：15，自引：0，他引：15
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。 相似文献
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牛型分枝杆菌MPB70蛋白胶乳凝集方法的建立及应用 总被引：9，自引：0，他引：9
为了研究牛结核病新型诊断试剂，提高诊断的特异性和敏感性，我们用大肠杆菌工程菌表达了牛型分枝杆菌特异性抗原MPB70并提纯蛋白建立了牛结核病乳胶凝集试验（LAT）诊断方法。MPB70是一种牛型分枝杆菌特异性分泌而卡介苗BCG缺失的蛋白质，热稳定性好，用此蛋白建立的乳胶凝集方法具有良好的敏感性、特异性以及较长的保存期，检测70份临床奶牛血清，与皮内变态反应和间接血凝方法相比较分别具有71．4％和88．6％的符合率。该方法还可鉴别诊断自然感染和疫苗免疫接种。我们建立的牛型分枝杆菌MPB70蛋白乳胶凝集试验诊断方法将分子生物学手段和经典试验方法有机结合，为临床快速检测牛结核病血清特异性抗体水平提供了行之有效的方法。 相似文献