[ Objective] To establish an indirect ELISA method which can detect fowl adenovirus group I (FAVI) antibody easily and rapidly. [ Method] The expressed and purified FAVI penton recombinant protein was used to be an ...[ Objective] To establish an indirect ELISA method which can detect fowl adenovirus group I (FAVI) antibody easily and rapidly. [ Method] The expressed and purified FAVI penton recombinant protein was used to be an antigen, optimized the reaction conditions, and then estab- lished the FAVI indirect penton-ELISA antibody detection method. [ Result] The optimal coating concentration of antigen was 1.5 μg/hole, the opti- mal coating condition was 37℃ 2 h and 4 ℃ overnight; the optimal dilution of serum was 1:100; the optimal working concentration of anti-chicken IgG-HRP was 1:2 000; the positive and negative critical value of ELISA was 0.335. Detected the 100 chicken serum samples by the established penton-ELISA method, the positive rate was 41%. [ Conclusion] Through the study, ~e established penton-ELISA method has a good specificity, sensitivity and reproducibility. And it offers an effective tool for the diagnosis of FAVI, the survey of antibody and epidemiology survey.展开更多
[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus s...[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus sp and Marteilia refringens from gene bank, design two pairs of spe- cific primers and two TaqMan probes with different fluorophores labeled. Optimizing the reactive conditions and reagent concentration in order that establishing the duplex real-time PCR method for detecting Perkinsus sp and Marteilia refringens simultaneously. [ Result ] The sensitivity of the du- plex real-time PCR method which about Pertdnsus sp and Marteilia refringens is 40 template copies. After combine the templates of Perkinsus sp and Marteilia refringens with different concentrations, this method still could be detect this two protozoan efficiently and synchronously. [ Condudon] The es- tablished duplex real-time PCR method for detecting Perkinsus sp. and Marteilia refringens possesses lots of advantages, such as specific, sensitive, rapid, quantitative and reproducible, can be used for clinical detection of infection which was caused by Perkinsus sp. and Marteilia refringens.展开更多
[ Objective] The aim was to explore novel method for treatment of Avian Reovirus. [ Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned in...[ Objective] The aim was to explore novel method for treatment of Avian Reovirus. [ Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned into the shRNA expression vector, namely, pSilencer-CMV 4.1 neo. Short hairpin RNA vector C1, C2, C3, which contain σC gene, and shRNA vector NS1, NS2, NS3, which contain aNS gene, were constructed separately. The constructed shRNA vectors and negative control were co-transfected into DF-1 cells with the eukaryotic expression vector pEG- FP-σC and pEGFP-σNS, respectively. [ Result] Observation through fluorescence microscope indicated that the constructed 6 shRNA could inhibit the expression of fusion protein to different degrees. In addition, results of Real-time PCR suggested that C3 and NS1 have the best interference effect to the viral duplication in vitro. [ Conclusionl Construction and selection of specific shRNA expression vectors inhibiting Avian Reovirus are significant for researching effects of σC and oNS proteins in infection and duplication of ARV, providing new idea for ARV antiviral therapy.展开更多
基金Guangxi Expert Special Fund Project (2011B020)Guangxi Science and Technology Research (0815009-3-6 and 10100014-5 )Guangxi Natural Science Foundation (2010GXNSFA013090)
文摘[ Objective] To establish an indirect ELISA method which can detect fowl adenovirus group I (FAVI) antibody easily and rapidly. [ Method] The expressed and purified FAVI penton recombinant protein was used to be an antigen, optimized the reaction conditions, and then estab- lished the FAVI indirect penton-ELISA antibody detection method. [ Result] The optimal coating concentration of antigen was 1.5 μg/hole, the opti- mal coating condition was 37℃ 2 h and 4 ℃ overnight; the optimal dilution of serum was 1:100; the optimal working concentration of anti-chicken IgG-HRP was 1:2 000; the positive and negative critical value of ELISA was 0.335. Detected the 100 chicken serum samples by the established penton-ELISA method, the positive rate was 41%. [ Conclusion] Through the study, ~e established penton-ELISA method has a good specificity, sensitivity and reproducibility. And it offers an effective tool for the diagnosis of FAVI, the survey of antibody and epidemiology survey.
文摘[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus sp and Marteilia refringens from gene bank, design two pairs of spe- cific primers and two TaqMan probes with different fluorophores labeled. Optimizing the reactive conditions and reagent concentration in order that establishing the duplex real-time PCR method for detecting Perkinsus sp and Marteilia refringens simultaneously. [ Result ] The sensitivity of the du- plex real-time PCR method which about Pertdnsus sp and Marteilia refringens is 40 template copies. After combine the templates of Perkinsus sp and Marteilia refringens with different concentrations, this method still could be detect this two protozoan efficiently and synchronously. [ Condudon] The es- tablished duplex real-time PCR method for detecting Perkinsus sp. and Marteilia refringens possesses lots of advantages, such as specific, sensitive, rapid, quantitative and reproducible, can be used for clinical detection of infection which was caused by Perkinsus sp. and Marteilia refringens.
基金supported by National Natural Science Foundation (31160512)Funds for Special Expert of Guangxi Province (2011B020)Guangxi Science and Techology Research Projects (0991222)
文摘[ Objective] The aim was to explore novel method for treatment of Avian Reovirus. [ Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned into the shRNA expression vector, namely, pSilencer-CMV 4.1 neo. Short hairpin RNA vector C1, C2, C3, which contain σC gene, and shRNA vector NS1, NS2, NS3, which contain aNS gene, were constructed separately. The constructed shRNA vectors and negative control were co-transfected into DF-1 cells with the eukaryotic expression vector pEG- FP-σC and pEGFP-σNS, respectively. [ Result] Observation through fluorescence microscope indicated that the constructed 6 shRNA could inhibit the expression of fusion protein to different degrees. In addition, results of Real-time PCR suggested that C3 and NS1 have the best interference effect to the viral duplication in vitro. [ Conclusionl Construction and selection of specific shRNA expression vectors inhibiting Avian Reovirus are significant for researching effects of σC and oNS proteins in infection and duplication of ARV, providing new idea for ARV antiviral therapy.