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1.
本文以试剂盒测血清β-羟丁酸含量大于1.2mmol/L作为亚临床酮病奶牛的诊断标准,采用酮粉法、试剂法、ROSS法和试纸法分别对血酮、乳酮和尿酮含量进行检测,并将定性检测的结果与定量检测的结果相比较。结果表明,酮粉法测血酮的符合率最高、为88.54%;试纸法测尿酮次之、为87.50%;ROSS法测尿酮的符合率和阳性率最低,分别为79.17%和13.54%。酮粉法测血酮敏感性最高,为70.37%;ROSS法测尿酮的敏感性最低,为37.04%;试剂法测乳酮、ROSS法测尿酮和试纸法测尿酮三者的特异性均为95.65%,高于酮粉法测血酮的特异性(94.20%),酮粉法测尿酮特异性最低、仅为88.41%。从操作难易度和成本上考虑,酮粉法测定乳酮含量比较适合牧场亚临床酮病的检测。 相似文献
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快速检测猪繁殖与呼吸综合征病毒胶体金免疫层析方法的建立 总被引：2，自引：0，他引：2
为建立一种快速、简便、灵敏的检测猪繁殖与呼吸综合征病毒(PRRSV)的胶体金免疫层析方法(GICA),本研究采用柠檬酸三钠还原法制备了胶体金颗粒,标记抗PRRSVN蛋白的单克隆抗体(MAb) 2D7制备免疫检测探针,将抗PRRSVN蛋白的MAb 1G7和羊抗鼠IgG抗体印迹在硝酸纤维素膜上,分别作为检测线和质控线,经条件优化,组装成胶体金免疫层析试纸条.本研究制备的PRRSV胶体金试纸条的最低检测限度为103.0 TCID50/mL;在特异性试验中,试纸条检测PRRSV呈阳性,其它主要猪病病原均为阴性;不同批次试纸条重复检测,结果无差异;对现地猪场送检的150份病料进行PRRSV病原检测,与RT-PCR相比较,试纸条的特异性和敏感性分别为98.13％和88.37％.两种方法的一致性Kappa值为0.882.建立的PRRSV抗原胶体金免疫层析检测方法具有良好的的敏感性、特异性、重复性及现地应用性.该试纸条的研制为PRRS的快速诊断及免疫预防提供了技术手段. 相似文献
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为了解不同猪口蹄疫O 型抗体检测试纸条的性能,通过检测已知相关及非相关阳性血清样品、阴性血清样品和免疫
猪血清样品,对4种猪口蹄疫O型抗体检测试纸条的敏感性、特异性进行研究评估,并对比猪口蹄疫O型VP1结构蛋白抗体ELISA诊断试剂盒的检测结果。结果表明,仅有A试纸条的敏感性和特异性较好,且与试剂盒抗体检测结果的符合率较高（90%）;而其他试纸条的敏感性和特异性较差,与试剂盒抗体检测结果的符合率较低（40%～65%）。通过对比检测,仅有A试纸条可用于大量临床样品的快速检测和现场检测,更适合基层兽医和养殖户使用,而其他试纸条的检测性能还有待进一步优化提升。 相似文献
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为研究虎红平板凝集试验(RBT)、全乳环状试验(MRT)、间接酶联免疫吸附试验(iELISA)及胶体金检测试纸条对布病的检测效果,本研究采用以上方法分别对感染地区和净化地区的牛群血清、羊群血清、牛群奶样进行了检测效果的比对。结果显示,针对牛群血清,RBT结果阳性率最高,iELISA与胶体金结果阳性率相似,表明RBT的敏感性较好,iELISA与胶体金检测试纸条的特异性较RBT好;针对羊群血清,iELISA与胶体金检测试纸条的敏感性和特异性较RBT好;针对牛群奶样,MRT结果阳性率低于全乳iELISA和胶体金检测试纸条。全乳iELISA和胶体金检测试纸条在试验中显示出具有较高的敏感性和特异性。 相似文献
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本试验利用胶体金免疫层析技术原理,研制了布鲁氏菌抗体检测试纸条,并以布鲁氏菌阳性血清国家标准品进行了敏感性试验,确定了最低检出量,同时与有可能存在交叉反应的血清和阴性血清进行了特异性试验;然后将保存不同时间的试纸条在进行了敏感性和特异性试验,证明其稳定期可以达到15个月。选择临床牛羊血清30份,利用该试纸条与布鲁氏菌病虎红平板试验抗原进行同步检测,发现两种试剂的检测符合率为98%。试验证明,所研制的布鲁氏菌抗体检测试纸条敏感性高、特异性强、稳定期长。 相似文献
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猪旋毛虫病快速检测试纸条的研制及特性测定 总被引：8，自引：0，他引：8
以旋毛虫重组分泌(excreting secretion．ES)抗原为基础,采用免疫金技术成功研制出旋毛虫病快速检测试纸条,试验结果显示试纸条与猪肺线虫、猪囊尾蚴、猪细颈囊尾蚴、猪米氏住肉孢子虫、猪蛔虫无交叉反应。证明旋毛虫病试纸条具有良好的特异性和敏感性,其敏感性与ELISA相当,与ELISA及消化法的检测结果符合率为100％。5min内肉眼观察获得结果,具有特异、敏感、快速、简单等特点,不需要任何专业技能和其他试剂。本方法的建立对猪旋毛虫病诊断和流行病学调查具有很大的应用价值。 相似文献
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《中国兽医学报》2020,(2):336-338
为了比较国产布鲁菌病间接ELISA抗体检测试剂盒、竞争ELISA抗体检测试剂盒和胶体金试纸条的检测效果,通过对已知阴阳性血清样品的检测,比较了上述3种检测方法的敏感性和特异性。进一步采用临床血清样品比较了3种检测方法与虎红平板凝集试验(RBT)的检测效果,并采用补体结合试验(CFT)对结果进行了复核。结果显示,间接ELISA、竞争ELISA和胶体金试纸条的敏感性分别为96.67%,100.00%和98.33%,3种检测方法的特异性分别为98.33%,93.33%和93.33%。对300份临床样品的检测结果显示,4种检测方法共同检出的阳性样本为27份,阴性样品为210份,整体符合率为79%。通过CFT对63份不同方法检验结果有差异的样本进行确诊,结果表明RBT与CFT的符合率最低,仅为3.17%;间接ELISA与CFT的符合率最高,为98.41%;竞争ELISA和胶体金试纸条的符合率分别为87.30%,85.71%。结果表明,间接ELISA、竞争ELISA和胶体金试纸条具有良好的特异性和敏感性,能够满足布病临床检测需求;RBT对临床样本的检测存在较高比例假阳性和假阴性。 相似文献
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通过制备脂多糖(LPS)并将其作为检测抗原,建立了一种检测奶样中布鲁氏菌抗体的酶联免疫层析方法。通过对处理方法和反应条件的优化,制备出敏感性高、特异性强的布鲁氏菌抗体检测试纸条。该试纸条敏感性为98.0%,抗体滴度为1:2~1:8,特异性为96.0%,且与其他常见病原无交叉反应,批内、批间重复性良好。保存期加速试验显示,该试纸条在2~30℃下可保存12个月;符合率验证显示,该方法与标准奶样的符合率达94.9%。上述结果表明,该检测方法具有敏感性高、特异性强以及方便、快捷等优点,适用于现场大批量检测,因而可应用于奶牛场布鲁氏菌病的快速诊断。 相似文献
11.
以基因工程表达的非洲猪瘟病毒VP73蛋白作为包被抗原,建立了间接ELISA方法,用以检测猪血清中抗非洲猪瘟VP73蛋白的抗体。该方法对非洲猪瘟标准阳性血清的检测灵敏度可以达到1∶2 560,与同类进口ELISA试剂盒相当。此方法只特异性检出非洲猪瘟阳性血清,而对猪传染性胸膜肺炎等5种猪传染病阳性血清的检测结果均为阴性,表明其具有良好的特异性。批内和批间重复性试验结果发现,检测同一份血清的变异系数小于10%,表明其重复性较好。包被好的酶标板37℃放置5d后,对同一份血清的检测敏感性无明显变化,初步表明其稳定性较好。利用建立的间接ELISA方法和进口ELISA试剂盒分别对150份血清样品进行非洲猪瘟血清抗体检测,结果表明本方法的特异性和敏感性分别为99.1%和94.3%,2种方法检测结果的符合率为98%。以上试验表明,本试验建立的间接ELISA方法具有良好的特异性和敏感性、较好的重复性和稳定性,可以满足临床检测的需求。 相似文献
12.
Confidence limits for prevalence of disease adjusted for estimated sensitivity and specificity 总被引：1，自引：0，他引：1
Prevalence of a disease is usually assessed by diagnostic tests that may produce false results. Rogan and Gladen (1978) described a method to estimate the true prevalence correcting for sensitivity and specificity of the diagnostic procedure, and Reiczigel et al. (2010) provided exact confidence intervals for the true prevalence assuming sensitivity and specificity were known. In this paper we propose a new method to construct approximate confidence intervals for the true prevalence when sensitivity and specificity are estimated from independent samples. To improve coverage we applied an adjustment similar to that described in Agresti and Coull (1998). According to an extensive simulation study the new confidence intervals maintain the nominal level fairly well even for sample sizes as small as 30; minimum coverage is above 88%, 93%, and 98% at nominal 90%, 95%, and 99%, respectively. We illustrate the advantages of the proposed method with real-life applications. 相似文献
13.
为了建立适用于基层养殖场快速、可视化的山羊痘病毒(GTPV)环介导等温扩增技术(LAMP)检测方法,根据GenBank公布的GTPV ORF103保守区核苷酸序列为靶序列,设计并合成一组特异性LAMP引物,优化反应条件(时间、温度)和反应体系(dNTP、内外引物浓度比),并进行特异性和灵敏度试验。结果表明,所建立的LAMP方法在65℃时反应50 min即可获得清晰的梯状条带;扩增物经10×SYBR GreenⅠ荧光染料染色,紫外线照射或日光下即可直接判定结果;灵敏度试验结果表明,该方法能够检出的GTPV核酸最低浓度为1.287 ng/mL,与常规PCR方法相比,灵敏度增高100倍;该方法特异性良好,与小反刍兽疫病毒(PPRV)、羊支原体(Mccp)、羊口疮病毒(ORFV)均无交叉反应;对10份临床样品检测结果显示,该方法与传统PCR检测结果一致。建立的LAMP检测方法可用于临床山羊痘病毒的检测。 相似文献
14.
Cloacal swabs were examined using a peroxidase-antiperoxidase (PAP) method that employs a monoclonal antibody to chlamydiae. The specificity and sensitivity of cloacal-swab PAP examination and the prevalence of chlamydiosis were calculated using both culture and tissue PAP evaluation as standards. The prevalence of chlamydial infections was 9.3% as determined by culture and 46.3% as judged by tissue PAP evaluation. Cloacal-swab PAP examination yielded a sensitivity of 71.4% and specificity of 85.5% when compared with culture results and a sensitivity of 81% and specificity of 85.7% compared with tissue PAP results. Culture had a sensitivity of 47.4% and specificity of 100% compared with tissue PAP results. Similarly, the efficiency of cloacal-swab PAP test was about 84% compared with culture results and tissue PAP. The efficiency of culture compared with tissue PAP was 73.1%. 相似文献
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Kvác M Kvetonová D Půzová G Ditrich O 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2003,50(8):405-411
This study involved the comparison of suitability of different methods for routine diagnostics of Cryptosporidium spp. Two staining methods, one concentration-sedimentation method, seven concentration-floatation methods and one combined floatation-sedimentation method were compared. The methods were tested with two concentrations (1 x 10(5) and 1 x 10(6)/g) of C. parvum and C. andersoni. The methods were evaluated using light microscope, magnification 400x for concentration methods and 1000x for stained samples respectively. Specificity of both staining methods was 95-100%. Ziehl-Neelsen with P < 0.01 is more suitable for identification of C. andersoni and modified Milácek-Vítovec with P < 0.01 for identification of C. parvum. Concerning specificity and sensitivity, the floatation-concentration method by Sheather was found to provide the best results of all selected methods. The merthiolate iodine formaldehyde concentration (MIFC) method was the least specific one. The least suitable method concerning sensitivity and costs was the floatation method with caesium chloride (CsCl) with a specificity of 29%. 相似文献
16.
试验旨在建立快捷、高效而准确的鉴别诊断猪德尔塔冠状病毒（Porcine deltacoronavirus,PDCoV）与传染性胃肠炎病毒（Transmissible gastroenteritis virus,TGEV）的双重实时荧光定量PCR方法。通过绘制双重实时荧光定量PCR的标准曲线,检验该方法的特异性、敏感性和重复性,并对临床样品进行检测。结果显示,双重实时荧光定量PCR方法的循环阈值与PDCoV和TGEV质粒拷贝数的对数之间存在良好的线性关系,且对应的相关系数分别为R（P）2=0.9994和R（T）2=0.996;能特异性地检测PDCoV和TGEV,而与PEDV、PRV、PRRSV、CSFV和RV无交叉反应,具有较强的特异性;检验PDCoV与TGEV质粒标准品的最低检测限度分别达到2和20拷贝/μL,且分别比常规RT-PCR高1 000和100倍,具有较高的敏感度;PDCoV与TGEV的批内和批间重复性检测的Ct均值基本相同,且变异系数（CV）均<2%,具有较好的重复性。用该方法对114份仔猪腹泻样品检测结果显示,PDCoV和TGEV的阳性率分别为5.6%（6/114）和8.8%（10/114）,混合感染检出率为4.6%（5/114）,比常规RT-PCR具有更高的检出率和敏感性。结果表明,本试验建立的双重实时荧光定量PCR方法具有特异性强、灵敏度高、重复性和稳定性好等优点,适用于病毒早期诊断和批量临床样品检测,为疾病防控、流行病学调查及相关性研究提供了技术支持及数据参考。 相似文献
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旨在建立检测非洲猪瘟病毒(ASFV)血清抗体的间接ELISA方法。本研究以两株纯化的ASFV p30与p54蛋白单克隆抗体(mAbs)为靶分子,利用噬菌体展示十二肽库进行四轮生物淘选,筛选多肽表位,以氨基酸GGG为接头设计合成表位串联多肽作为包被抗原,通过棋盘滴定法确定间接ELISA的最佳反应条件,利用不同类型血清样本对建立的方法进行特异性分析、敏感度分析、稳定性分析及符合性评价。噬菌体淘选试验结果表明146PAEPYTT152为本实验室保存的mAb所识别的p54蛋白抗原表位核心序列。ELISA条件优化试验结果显示,以鸡卵白蛋白(OVA)作为N端偶联物的表位串联多肽抗原具有较低的非特异性血清反应背景,当多肽以碳酸盐缓冲液包被(2 μg·mL-1),血清以封闭液(1%明胶溶液)稀释100倍,辣根过氧化物酶(HRP)标记抗体以0.05% PBST溶液稀释5 000倍时,反应效果最佳;以上述优化后的条件确定了血清抗体阳性临界值为0.339。方法评价试验结果显示,该方法与经典猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)、猪细小病毒(PPV)及猪伪狂犬病病毒(PRV)的抗体阳性血清均无交叉反应,能检测低至1 600倍稀释的ASFV阳性血清,具有较好的重复性。用该方法与商品化的ASFV抗体检测试剂盒同时检测320份猪血清样本,两种方法的相对特异性和相对敏感性分别为97.6%与97.3%,总体符合率达97.5%(312/320)。综上表明,本研究建立的多肽间接ELISA方法具有良好的特异性、敏感性及重复性,具有发展为临床诊断试剂盒的潜在应用价值。 相似文献
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为建立更敏感、特异的牛片形吸虫病早期诊断方法,本试验将收集的大片吸虫分泌排泄产物（Fasciola gigantica excretory-secretory products,FgESP）进行凝胶过滤层析并从中筛选出检测效果最优的UV280吸收峰,从此峰对应的组分中筛选出诊断效果最优的层析组分后将其作为抗原,通过棋盘滴定试验优化抗原包被浓度、血清稀释度、酶标二抗稀释度、显色时间等,确定临界值,建立牛片形吸虫病间接ELISA诊断方法。对建立的方法进行敏感性和特异性试验,检测试验感染0~14周的水牛血清和临床160份水牛血清样本,并与已有的诊断抗原FgESP（阴阳临界值为0.320）进行诊断效果比较。结果显示,层析组分F22具有较优的检测效果。间接ELISA的最佳反应条件为：F22抗原包被浓度0.157 μg/mL,待测血清与酶标二抗的稀释度分别为1：400和1：40 000,显色时间为25 min。应用建立的方法检测15份水牛阴性血清,确定D450 nm阴阳性临界值为0.408。比较F22与FgESP发现,二者特异性相似,但F22作为抗原的敏感性高于FgESP。检测试验感染大片吸虫的水牛0~14周血清,结果表明,从感染第2周开始F22-IgG水平极显著高于FgESP-IgG水平（P<0.01）,因此,F22比FgESP诊断效果更优。对160份水牛血清检测发现,F22阳性检出率为71.25%,FgESP阳性检出率为63.75%。上述结果表明,相比于FgESP,以F22作为诊断抗原建立的大片吸虫病间接ELISA诊断方法敏感性更高,可更有效地进行牛大片吸虫病的早期诊断。 相似文献
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Serodiagnosis of ovine paratuberculosis, using lipoarabinomannan in an enzyme-linked immunosorbent assay 总被引：7，自引：0，他引：7
E A Sugden A H Corner B S Samagh B W Brooks C Turcotte K H Nielsen R B Stewart J R Duncan 《American journal of veterinary research》1989,50(6):850-854
The use of lipoarabinomannan (LAM; obtained from Mycobacterium paratuberculosis) in and ELISA (LAM-ELISA) to test 75 sheep sera from a paratuberculosis-infected flock resulted in an approximate threefold increase in sensitivity (from 23.5% to 70.6%), compared with the use of Annau's polysaccharide in a complement fixation test (P-CFT). Even after manipulation of the LAM-ELISA cut-off value to produce a specificity of 100% to match that of the P-CFT, the sensitivity still was approximately twofold greater than that of the P-CFT. Anti-bovine monoclonal antiglobulin-enzyme conjugates matched commercially available anti-ovine polyclonal antiglobulin-enzyme conjugates with respect to sensitivity and specificity. False-positive results were found to be less frequent after combining 2 serodiagnostic tests, LAM-ELISA and D antigenagar gel immunodiffusion, resulting in an increase in specificity from 88.1% to 95.2%. The repeatability of true seropositive and seronegative results was found to be 89.5% and 91.1%, respectively, for sera obtained less than or equal to 1 month prior to slaughter and 91.7% and 95.5%, respectively, for reanalysis of sera obtained at the time of slaughter. 相似文献
20.
Latent-class models were used to determine the sensitivity, specificity and predictive values of a polyclonal blocking enzyme-linked immunosorbent assay (ELISA) and a modified complement-fixation test (CFT) when there was no reference test. The tests were used for detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in a survey of respiratory diseases in Danish finishing pigs. The estimates were obtained by maximum-likelihood and also by a Bayesian method (implemented with Gibbs sampling). Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence, given the true disease status.
No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity and specificity of the ELISA were 100% and 92.8% (90.1–95.5%) and the corresponding values of the CFT were 90.6% (85.8–95.4%) and 98.6% (98.0–99.3%), respectively. Bayesian estimates and posterior 95% credible intervals of the sensitivity and specificity of the ELISA were 99.7% (98.7–100%) and 92.7% (89.9–95.3%) and of the CFT were 90.6% (86.0–95.3%) and 98.7% (98.0–99.3%). The sensitivity and specificity of a combined test, where the CFT is subsequently applied to the pig sera that test positive in the ELISA, were estimated at 90.2% (85.6–95.0%) and 99.9% (99.8–100%), respectively. The cost of the combined test was less than the cost of the use of the CFT alone, at prevalences <54%. Prevalences and predictive values and their 95% limits were estimated in six sub-samples of data. The estimates of sensitivity and specificity obtained in the present investigation generally validate those reported from other sources. 相似文献