为给猫传染性腹膜炎病毒(FIPV)的流行和地理分布提供有价值的检测手段,以FIPV S蛋白为包被抗原,利用FIPV的特异性单克隆抗体(FIPV MAb 3D8)为竞争抗体,经条件优化,建立一种快速检测FIPV抗体的竞争ELISA法。优化结果表明最佳条件为:包被...为给猫传染性腹膜炎病毒(FIPV)的流行和地理分布提供有价值的检测手段,以FIPV S蛋白为包被抗原,利用FIPV的特异性单克隆抗体(FIPV MAb 3D8)为竞争抗体,经条件优化,建立一种快速检测FIPV抗体的竞争ELISA法。优化结果表明最佳条件为:包被抗原质量浓度10μg/mL,血清稀释度1∶4;FIPV MAb 3D8稀释倍数1∶100,酶标二抗稀释倍数1∶5000;CBS在37℃包被1 h,5%BSA在37℃封闭2 h,FIPV MAb 3D8在37℃孵育1 h,TMD底物显色液37℃避光5 min。临界值PI≥25.76%时为阳性,PI<20.22%为阴性。该方法特异性较强,重复性较好。利用该方法和已发表的其他方法同时对来自宠物医院的62份临床血清样本进行检测,总体符合率为85.5%,表明该方法准确性较高。本研究建立了快速准确测定FIPV抗体的竞争ELISA方法,适用于临床样本检测,为猫传染性腹膜炎的鉴定、防控和流行病学调查提供支持。展开更多
Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was ...Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.展开更多
Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect ...Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO. The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively. The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively.展开更多
Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monocl...Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.展开更多
文摘为给猫传染性腹膜炎病毒(FIPV)的流行和地理分布提供有价值的检测手段,以FIPV S蛋白为包被抗原,利用FIPV的特异性单克隆抗体(FIPV MAb 3D8)为竞争抗体,经条件优化,建立一种快速检测FIPV抗体的竞争ELISA法。优化结果表明最佳条件为:包被抗原质量浓度10μg/mL,血清稀释度1∶4;FIPV MAb 3D8稀释倍数1∶100,酶标二抗稀释倍数1∶5000;CBS在37℃包被1 h,5%BSA在37℃封闭2 h,FIPV MAb 3D8在37℃孵育1 h,TMD底物显色液37℃避光5 min。临界值PI≥25.76%时为阳性,PI<20.22%为阴性。该方法特异性较强,重复性较好。利用该方法和已发表的其他方法同时对来自宠物医院的62份临床血清样本进行检测,总体符合率为85.5%,表明该方法准确性较高。本研究建立了快速准确测定FIPV抗体的竞争ELISA方法,适用于临床样本检测,为猫传染性腹膜炎的鉴定、防控和流行病学调查提供支持。
基金Supported by National High-tech R&D Program (863) Subsidized Project(2006AA10A204)Special Fund for Basic Scientific Research-related Subsidy of State-level and Public-welfare Scientific Research Institutes~~
文摘Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.
文摘Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO. The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively. The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively.
基金Supported by the Key Project of National Science and Technology Surporting Plan during 11th-Five-Year of China(2006BAK02A21/1)~~
文摘Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.
