基于上轉(zhuǎn)發(fā)光免疫層析技術(shù)的常見食源性致病菌快速檢測(cè)方法研究與評(píng)價(jià)
發(fā)布時(shí)間:2018-12-06 17:55
【摘要】:目的研制針對(duì)4種常見食源性致病菌:甲型副傷寒沙門菌(Salmonella paratyphi A)、乙型副傷寒沙門菌(Salmonella paratyphi B)、大腸埃希菌O157∶H7(Escherichia coli O157∶H7)、副溶血弧菌(Vibrio parahaemolyticus)的基于上轉(zhuǎn)發(fā)光免疫層析技術(shù)(UPT-LF)的快速定量檢測(cè)試紙,并對(duì)其檢測(cè)性能進(jìn)行評(píng)價(jià)。方法以上轉(zhuǎn)發(fā)光納米顆粒(UCP-NPs)作為示蹤物,基于雙抗體夾心檢測(cè)模型,研制針對(duì)上述4種靶標(biāo)菌的UPT-LF試紙;以4種靶標(biāo)菌的系列濃度標(biāo)準(zhǔn)菌懸液作為標(biāo)準(zhǔn)樣品,評(píng)價(jià)試紙的敏感性、特異性、線性和精密性,并進(jìn)行模擬染菌食品的檢測(cè),評(píng)價(jià)模擬樣品陽(yáng)性檢出率。結(jié)果針對(duì)4種靶標(biāo)菌的UPT-LF試紙均具有良好的線性,相關(guān)系數(shù)r為0.985~0.996;檢測(cè)敏感性達(dá)105~106CFU/ml,與其他近緣菌株無(wú)交叉反應(yīng);對(duì)乳制品、飲料、小食品、水產(chǎn)、肉類等細(xì)菌污染的食品檢出率較高。結(jié)論該研究建立的對(duì)4種常見食源性致病菌進(jìn)行快速檢測(cè)的UPT-LF,簡(jiǎn)便快速,具有良好的敏感性、特異性和線性定量能力,操作性能可滿足食品安全檢測(cè)的要求。
[Abstract]:Objective to develop four kinds of common foodborne pathogenic bacteria: Salmonella paratyphi A (Salmonella paratyphi A), Salmonella paratyphi B (Salmonella paratyphi B), Escherichia coli (O157:H7 (Escherichia coli O157:H7). Rapid quantitative detection of Vibrio parahaemolyticus (Vibrio parahaemolyticus) based on up-conversion luminescence immunochromatography (UPT-LF) was carried out and its detection performance was evaluated. Methods the above forwarded light nanoparticles (UCP-NPs) were used as tracers. Based on the double antibody sandwich detection model, the UPT-LF test paper for the above four target bacteria was prepared. The sensitivity, specificity, linearity and precision of the test paper were evaluated by using the suspension of four kinds of target bacteria as the standard sample, and the positive rate of the simulated sample was evaluated. Results the UPT-LF test paper for the four target bacteria had a good linearity with a correlation coefficient of 0.996, a sensitivity of 105 ~ 106CFU / ml, and no cross reaction with other close strains. For dairy products, beverages, small foods, aquatic products, meat and other bacterial contamination of food detection rate is higher. Conclusion the UPT-LF, developed in this study for the rapid detection of four common foodborne pathogenic bacteria is simple and rapid, has good sensitivity, specificity and linear quantitative ability, and the operational performance can meet the requirements of food safety detection.
【作者單位】: 解放軍總醫(yī)院臨床檢驗(yàn)科;軍事醫(yī)學(xué)科學(xué)院微生物流行病研究所病原微生物生物安全國(guó)家重點(diǎn)實(shí)驗(yàn)室;海軍總醫(yī)院內(nèi)分泌科;重慶醫(yī)科大學(xué)公共衛(wèi)生與管理學(xué)院;
【基金】:國(guó)家863計(jì)劃資助項(xiàng)目(2013AA032205) 國(guó)家科技重大專項(xiàng)資助項(xiàng)目(2011ZX10004-001,2012ZX10004801-002-004,2012ZX10004801-004-015) 北京市科委2014年度科技創(chuàng)新基地培育與發(fā)展工程專項(xiàng)資助項(xiàng)目(Z141107004414026) 國(guó)家自然科學(xué)基金資助項(xiàng)目(81000774) 北京市科技新星資助項(xiàng)目
【分類號(hào)】:R155.5
[Abstract]:Objective to develop four kinds of common foodborne pathogenic bacteria: Salmonella paratyphi A (Salmonella paratyphi A), Salmonella paratyphi B (Salmonella paratyphi B), Escherichia coli (O157:H7 (Escherichia coli O157:H7). Rapid quantitative detection of Vibrio parahaemolyticus (Vibrio parahaemolyticus) based on up-conversion luminescence immunochromatography (UPT-LF) was carried out and its detection performance was evaluated. Methods the above forwarded light nanoparticles (UCP-NPs) were used as tracers. Based on the double antibody sandwich detection model, the UPT-LF test paper for the above four target bacteria was prepared. The sensitivity, specificity, linearity and precision of the test paper were evaluated by using the suspension of four kinds of target bacteria as the standard sample, and the positive rate of the simulated sample was evaluated. Results the UPT-LF test paper for the four target bacteria had a good linearity with a correlation coefficient of 0.996, a sensitivity of 105 ~ 106CFU / ml, and no cross reaction with other close strains. For dairy products, beverages, small foods, aquatic products, meat and other bacterial contamination of food detection rate is higher. Conclusion the UPT-LF, developed in this study for the rapid detection of four common foodborne pathogenic bacteria is simple and rapid, has good sensitivity, specificity and linear quantitative ability, and the operational performance can meet the requirements of food safety detection.
【作者單位】: 解放軍總醫(yī)院臨床檢驗(yàn)科;軍事醫(yī)學(xué)科學(xué)院微生物流行病研究所病原微生物生物安全國(guó)家重點(diǎn)實(shí)驗(yàn)室;海軍總醫(yī)院內(nèi)分泌科;重慶醫(yī)科大學(xué)公共衛(wèi)生與管理學(xué)院;
【基金】:國(guó)家863計(jì)劃資助項(xiàng)目(2013AA032205) 國(guó)家科技重大專項(xiàng)資助項(xiàng)目(2011ZX10004-001,2012ZX10004801-002-004,2012ZX10004801-004-015) 北京市科委2014年度科技創(chuàng)新基地培育與發(fā)展工程專項(xiàng)資助項(xiàng)目(Z141107004414026) 國(guó)家自然科學(xué)基金資助項(xiàng)目(81000774) 北京市科技新星資助項(xiàng)目
【分類號(hào)】:R155.5
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