【摘要】：建立了生物素-鏈霉親和素時(shí)間分辨熒光免疫分析(BAS-TRFIA)高靈敏檢測(cè)嗜水氣單胞菌的新方法.以兔抗IgG包被微孔板,加入嗜水氣單胞菌菌株B18和生物素化IgG,生成的夾心復(fù)合物用Eu~(3+)-鏈霉親和素作為示蹤物,Eu~(3+)可與離解增強(qiáng)液形成時(shí)間分辨熒光(TRF),通過(guò)TRF值對(duì)病原菌定量.方法的檢測(cè)限為1.0×10~2 cfu/mL,在1.0×10~1.0×10~6 cfu/mL范圍內(nèi)線(xiàn)性良好,相關(guān)系數(shù)0.9716.用該法檢測(cè)時(shí),不同來(lái)源的嗜水氣單胞菌以及其它氣單胞菌均呈陰性.板內(nèi)和板間的變異系數(shù)分別小于5.00%和9.00%.所有相關(guān)檢測(cè)試劑37℃恒溫放置6 d后,檢測(cè)性能無(wú)明顯改變.對(duì)60份人工感染的美洲鰻鱺組織樣品,包括肝、腎、腸、鰓和肌肉等組織進(jìn)行檢測(cè),陽(yáng)性檢測(cè)率為98.33%.結(jié)果表明,BAS-TRFIA法檢測(cè)嗜水氣單胞菌,靈敏度高、特異性好、操作簡(jiǎn)便,該方法為水產(chǎn)養(yǎng)殖病原菌檢測(cè)提供了新的思路.
[Abstract]:A highly sensitive method for the detection of Aeromonas hydrophila by biotin-streptavidin time resolved fluorescence immunoassay (BAS-TRFIA) was established. The micropore plate of rabbit anti IgG was coated with B18 strain of Aeromonas hydrophila and biotinylated IgG. The sandwich complex was produced with EU3-Streptomyces avidin as a tracer, which could be used to form a time resolved fluorescent (TRF), with dissociation enhancer to quantify the pathogenic bacteria by TRF value. The detection limit of the method is 1.0 脳 10 ~ (-2) cfur 路mL ~ (-1), the linear range is 1.0 脳 10 ~ (10) 脳 10 ~ (-6) cfu/mL, the correlation coefficient is 0.9716. When detected by this method, Aeromonas hydrophila and other Aeromonas from different sources were negative. The coefficient of variation within and between boards was less than 5.00% and 9.00%, respectively. The detection performance of all the related reagents remained unchanged after 6 days at 37 鈩，

本文編號(hào)：2139350