A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluore...A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in mi- cro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H2O2)-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL·1. The detection limit of CEA was 0.61 ng mL·1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation.展开更多
Banana streak virus (BSV) and Sugarcane bacilliform virus (SCBV) are two badnaviruses commonly found in all banana growing areas of the world. It is a threat to the production and improvement of Musa germplasm. In Bur...Banana streak virus (BSV) and Sugarcane bacilliform virus (SCBV) are two badnaviruses commonly found in all banana growing areas of the world. It is a threat to the production and improvement of Musa germplasm. In Burkina Faso, the presence of badnaviruses was reported in banana producing regions. The objective of this study was to determine the prevalence of BSV and SCBV in banana production areas of Burkina Faso. A survey followed by a symptomatologic study was conducted in banana plantations in 27 localities of the nine main banana producing regions from July to October 2018 and September to December 2020. In all, 251 leaf samples were collected and analysed for BSV and SCBV infection by Indirect Antigen Coated Plate Assay-ELISA followed by amplification of the RT/RNase H region using Polymerase chain reaction with Badna FP/RP and SCBV F/R primers, respectively. A variety of symptoms were observed on almost all plant organs which were revealed due to BSV by symptomatologic study. The results of serological and molecular diagnosis revealed a high overall prevalence of BSV in 80.48% of the samples tested. BSV was distributed in seven survey regions out of nine with prevalence ranging from 10% to 100% in North, Centre, Centre West, Hauts Bassins, Cascades, Centre East and Boucle of Mouhoun regions. Very low prevalence was recorded for SCBV in Cascades and East Centre region with 4.35 and 12.5%, respectively. Species detection using specific primers to each species revealed three main species: Banana streak Obino l’ewaï virus (BSOLV), Goldfinger virus (BSGFV) and Imové virus (BSIMV) in the samples tested, respectively in the proportions of 23%, 8% and 0.8%. Co-infection between BSV species was also detected.展开更多
Rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（...Rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（DE3） using the pMAL-C2 X expression vector. The recombinant protein was used as the immunogen to immunize BALB/c mice. A hybridoma cell line 8A12 secreting monoclonal antibody（MAb） against RRSV was obtained by fusing mouse myeloma cells（Sp 2/0） with spleen cells from the immunized BALB/c mice. Western blot analysis showed that the MAb 8A12 can specifically react with RRSV CP. Using the MAb, an antigen-coated-plate enzyme-linked immunosorbent assay（ACP-ELISA）, a dot enzyme-linked immunosorbent assay（dot-ELISA）, and immunocapture-RT-PCR（IC-RT-PCR） assay were developed to detect RRSV. The established ACP-ELISA, dot-blot ELISA and IC-RT-PCR methods could detect RRSV in infected rice tissue crude extracts with dilutions of 1：40 960, 1：1 280 and 1：655 360（w/v, g mL-1）, respectively. The ACP-ELISA and dot-blot ELISA methods could detect RRSV in infected insect vector crude extracts with dilutions of 1：12 800 and 1：1 600（an individual planthopper μL-1）, respectively. The field survey revealed that Rice ragged stunt disease occurs on rice in Hainan, Yunnan, Guangxi, Sichuan, Guizhou, Fujian, Hunan, Jiangxi and Zhejiang in China.展开更多
目的：观察CD31在带部分松质骨的小牛皮质骨板移植愈合过程中不同时相的表达及组织分布情况，探讨其愈合机制及再血管化情况与骨改建的关系，为进一步研究组织工程骨奠定实验基础。方法实验动物选用健康3月龄36只新西兰大白兔。植入带...目的：观察CD31在带部分松质骨的小牛皮质骨板移植愈合过程中不同时相的表达及组织分布情况，探讨其愈合机制及再血管化情况与骨改建的关系，为进一步研究组织工程骨奠定实验基础。方法实验动物选用健康3月龄36只新西兰大白兔。植入带部分松质骨的小牛皮质骨骨板，分别于术后4、8、12、24周各时间点取材，通过一般情况及大体标本、X线检查、组织学及CD31免疫组织化学染色观察其愈合情况，并借助计算机图像分析仪进行定量分析，分别用再血管化指数与新骨形成面积代表再血管化和骨改建程度。结果⑴X线显示：术后24周见明显愈合。⑵在移植骨愈合的不同阶段，CD31的表达存在不同。术后4周、8周，移植骨周围的软组织、结合部、周围的骨痂及软骨巢周围见较多强阳性表达；松质骨内较多阳性表达，内见单个、条索状、团簇状新生血管，移植骨内部扩大的哈弗氏管及周围见阳性表达；术后24周，移植骨内扩大的哈弗氏管及周围呈强阳性表达，新生血管继续增多。⑶对新骨形成面积和再血管化指数的计量资料进行相关性分析表明，再血管化与骨改建呈高度正相关（ r ＝0．984）。结论 CD31的阳性表达贯穿于异种骨愈合的整个过程，而且不同时段、不同部位的CD31表达是不同的，并可以反映移植骨的再血管化与骨改建状况。骨移植后再血管化程度与骨改建密切相关。展开更多
基金support from the CAS Major Scientific Research and Equipment Development Projects (YZ-0632)the National Natural Science Founolation of China (Grant No. 20907060)+2 种基金the CAS Graduate Innovation Foundation (No. YXLW-3)the National key Technology R&D Program (Grant No. 2006BAF07B03-1-1)Shaanxi Province, China’s Scholarship Council (Grant No. 08JK482)
文摘A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in mi- cro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H2O2)-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL·1. The detection limit of CEA was 0.61 ng mL·1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation.
文摘Banana streak virus (BSV) and Sugarcane bacilliform virus (SCBV) are two badnaviruses commonly found in all banana growing areas of the world. It is a threat to the production and improvement of Musa germplasm. In Burkina Faso, the presence of badnaviruses was reported in banana producing regions. The objective of this study was to determine the prevalence of BSV and SCBV in banana production areas of Burkina Faso. A survey followed by a symptomatologic study was conducted in banana plantations in 27 localities of the nine main banana producing regions from July to October 2018 and September to December 2020. In all, 251 leaf samples were collected and analysed for BSV and SCBV infection by Indirect Antigen Coated Plate Assay-ELISA followed by amplification of the RT/RNase H region using Polymerase chain reaction with Badna FP/RP and SCBV F/R primers, respectively. A variety of symptoms were observed on almost all plant organs which were revealed due to BSV by symptomatologic study. The results of serological and molecular diagnosis revealed a high overall prevalence of BSV in 80.48% of the samples tested. BSV was distributed in seven survey regions out of nine with prevalence ranging from 10% to 100% in North, Centre, Centre West, Hauts Bassins, Cascades, Centre East and Boucle of Mouhoun regions. Very low prevalence was recorded for SCBV in Cascades and East Centre region with 4.35 and 12.5%, respectively. Species detection using specific primers to each species revealed three main species: Banana streak Obino l’ewaï virus (BSOLV), Goldfinger virus (BSGFV) and Imové virus (BSIMV) in the samples tested, respectively in the proportions of 23%, 8% and 0.8%. Co-infection between BSV species was also detected.
基金supported by the National Basic Research Program of China(2010CB126203)the special fund for Agro-scientific Research in the Public Interest,China(201003031)+1 种基金Earmarked Funds for Modern Agro-industry Technology Research SystemZhejiang Provincial Natural Science Foundation of China(Z3090039)
文摘Rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（DE3） using the pMAL-C2 X expression vector. The recombinant protein was used as the immunogen to immunize BALB/c mice. A hybridoma cell line 8A12 secreting monoclonal antibody（MAb） against RRSV was obtained by fusing mouse myeloma cells（Sp 2/0） with spleen cells from the immunized BALB/c mice. Western blot analysis showed that the MAb 8A12 can specifically react with RRSV CP. Using the MAb, an antigen-coated-plate enzyme-linked immunosorbent assay（ACP-ELISA）, a dot enzyme-linked immunosorbent assay（dot-ELISA）, and immunocapture-RT-PCR（IC-RT-PCR） assay were developed to detect RRSV. The established ACP-ELISA, dot-blot ELISA and IC-RT-PCR methods could detect RRSV in infected rice tissue crude extracts with dilutions of 1：40 960, 1：1 280 and 1：655 360（w/v, g mL-1）, respectively. The ACP-ELISA and dot-blot ELISA methods could detect RRSV in infected insect vector crude extracts with dilutions of 1：12 800 and 1：1 600（an individual planthopper μL-1）, respectively. The field survey revealed that Rice ragged stunt disease occurs on rice in Hainan, Yunnan, Guangxi, Sichuan, Guizhou, Fujian, Hunan, Jiangxi and Zhejiang in China.
文摘目的：观察CD31在带部分松质骨的小牛皮质骨板移植愈合过程中不同时相的表达及组织分布情况，探讨其愈合机制及再血管化情况与骨改建的关系，为进一步研究组织工程骨奠定实验基础。方法实验动物选用健康3月龄36只新西兰大白兔。植入带部分松质骨的小牛皮质骨骨板，分别于术后4、8、12、24周各时间点取材，通过一般情况及大体标本、X线检查、组织学及CD31免疫组织化学染色观察其愈合情况，并借助计算机图像分析仪进行定量分析，分别用再血管化指数与新骨形成面积代表再血管化和骨改建程度。结果⑴X线显示：术后24周见明显愈合。⑵在移植骨愈合的不同阶段，CD31的表达存在不同。术后4周、8周，移植骨周围的软组织、结合部、周围的骨痂及软骨巢周围见较多强阳性表达；松质骨内较多阳性表达，内见单个、条索状、团簇状新生血管，移植骨内部扩大的哈弗氏管及周围见阳性表达；术后24周，移植骨内扩大的哈弗氏管及周围呈强阳性表达，新生血管继续增多。⑶对新骨形成面积和再血管化指数的计量资料进行相关性分析表明，再血管化与骨改建呈高度正相关（ r ＝0．984）。结论 CD31的阳性表达贯穿于异种骨愈合的整个过程，而且不同时段、不同部位的CD31表达是不同的，并可以反映移植骨的再血管化与骨改建状况。骨移植后再血管化程度与骨改建密切相关。
