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Chinese Science Bulletin 2011年第8期56卷 749-754页

作者： LI WeiHua MENG Meng he fangyang WAN YuPing XUE HuYin LIU Wei YIN WeiWei XU Jing FENG CaiWei WANG ShanLiang LU Xiao LIU JinTing XI RiMo School of Chemistry and Chemical Engineering Shandong University Jinan 250100 China College of Pharmacy and Tianjin key Laboratory of Molecular Drug Research Nankai University Tianjin 300071 China Beijing Wanger Biotechnology Co. Ltd. Beijing 102206 China Central Laboratory of Shandong Academy of Agricultural Sciences Jinan 250100 China

Based on the preparation of an anti-diethylstilbestrol(DES) monoclonal antibody,a simple and convenient indirect competitive enzyme-linked immunosorbent assay(ELISA) method for DES detection has been *** monoclonal antibody demonstrated high sensitivity to DES with an IC50 value of 275 pg mL-1 and detection limit(LOD) of 90 pg *** specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol(he developed method,and the recovery rates were greater than 79.5%.Intra-and inter-assay variations were about 6%.This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real *** LODs in fish/shrimp,liver,feed and urine spiked with DES were 600,600,4800 and 600 pg mL-1,*** results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals.

关键词： ELISA检测方法单克隆抗体己烯雌酚生物样品竞争间接酶联免疫吸附试验 DES

Study on Rapid Detection of Tetracyclines,Fluoroquinolones and Sulfonamides in Milk

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Agricultural Biotechnology 2019年第1期8卷 241-245页

作者： Yuping WAN Xiaosheng WU Tingting CUI Dongshan CAO Zhaoqin WANG Linchen WANG Chun-yan YANG fangyang he Beijing Kwinbon Biotechnology Co. Ltd. Beijing Engineering Research Center of Food Safety Immunodetection Beijing Wanger Biotechnology Co. Ltd.

Aiming at the market demand for rapid detection of tetracyclines,fluoroquinolones and sulfonamides in milk,a golloidal gold immunochromatography test strip for simultaneous detection of tetracyclines,fluoroquinolones and sulfonamides in milk was prepared based on the principle of competitive inhibition immunochromatography. The performance indicators of the test strip were verified. The results showed that the test strip can simultaneously detect 4 tetracyclines,13 fluoroquinolones and 13 sulfonamides,and the detection limits all can meet the national residue limits; the tests strip exhibited false positive rate≤5% and false negative rate = 0; and no cross-reaction with other drugs was commonly found in milk,indicating good specificity. The method is simple,rapid,and has low cost and easy popularization. It provides a means for realizing on-site rapid detection and is of important practical significance to guarantee of safety of milk and dairy products in China.

关键词： Milk Tetracyclines Fluoroquinolones Sulfonamides Colloidal gold immunochromatography assay

Study on Colloidal Gold Immunochromatography Assay for Rapid Detection of Spectinomycin

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Agricultural Biotechnology 2019年第1期8卷 188-189,193页

作者： Zhaoqin WANG Yuping WAN Xiaosheng WU Yu ZHANG Fangfang JIA Guangyao HAN Zhengxue PENG fangyang he Beijing Kwinbon Biotechnology Company Beijing Engineering Research Centre of Food Safety Immunodetection Beijing Wanger Biotechnology Company

[Objectives] This study was conducted to establish a rapid detection method for spectinomycin in pork,chicken,fish,shrimp flesh and water.[Methods]A test strip for rapid detection of spectinomycin in milk was developed by colloidal gold immunochromatography assay. [Results]The test strip had a detection limit of 50 μg/kg to milk with a detection time of 15 min,and the false positive rate and false negative rate were both 0. [Conclusions]The method is accurate,simple,reliable and convenient,and is suitable for rapid on-site spectinomycin detection.

关键词： Spectinomycin Gold immunochromatography assay rapid test strip Rapid detection

Study on the Production of Pentachloronitrobenzene Monoclonal Antibody and Its ELISA Kit for Rapid Detection

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Agricultural Biotechnology 2021年第5期10卷 17-20,44页

作者： Yuhua MA Kuo ZHANG Jianxiong ZHANG Fangfang JIA fangyang he Yanan CUI Mingyang LI Yuping WAN Beijing Kwinbon Biotechnology Company Beijing 102206China Beijing Engineering Research Centre of Food Safety Immunodetection Beijing 102206China Beijing Yanqing Animal Disease Prevention and Control Center Beijing 102100China Tangshan General Inspection and Testing Centre for Food and Drug Tangshan 063000China

[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in ***.[Results]The standard curve range of the kit was 0-8.1μg/L;the detection limit for *** was 0.912μg/kg;the recovery was 80.6%-103.5%;and the relative standard deviation range within batches was 5.3%-10.1%,and the relative standard deviation range between batches was 6.7%-8.1%.The specificity of the pentachloronitrobenzene monoclonal antibody was relatively good,and the cross-reaction rates with pentachlorophenol,hexachlorobenzene,tetrachlorophthalide,and chlorothalonil were low,all of which did not exceed 30%.The ELISA kit could be stored at 4℃for 12 months,showing good stability.[Conclusions]The detection kit has low cost,short time and small deviation,and is an ideal preliminary screening method.

关键词： Pentachloronitrobenzene Monoclonal antibody Enzyme linked immunosorbent assay kit

Colloidal Gold Test Strip for the Detection of Fluoroquinolones in Foods Using an Analyzer

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Agricultural Biotechnology 2019年第2期8卷 172-176页

作者： Fangfang JIA Yuping WAN Linchen WANG Xiping WANG Zhaoqin WANG fangyang he Haifeng CUI Jing LI Beijing Kwinbon Biotechnology Co. Ltd. Beijing Engineering Research Center of Food Safety Immunodetection Beijing Wanger Biotechnology Co. Ltd.

A test strip was developed to rapidly detect fluoroquinolones in foods by colloidal gold immunochromatography method in combination with a food safety analyzer. The results showed that the test strip has the detection sensitivity of 20 μg/L for enrofloxacin, sarafloxacin, difloxacin, ofloxacin, norfloxacin, ciprofloxacin, pefloxacin, flumequine and danofloxacin, and the detection sensitivity of 40 μg/L for enoxacin and oxolinic acid. The test strip consumes only 10 min for detection, and the false positive rate and the false negative rate are both zero. The test strip can accurately, reliably and easily detect residual fluoroquinolones in foods, and is suitable for on-site detection of a large number of samples.

关键词： Fluoroquinolones Colloidal gold strip Food safety analyzer Rapid detection

Development of a Kind of Immunomagnetic Bead Kit to Separate and Enrich Salbutamol

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Agricultural Biotechnology 2018年第6期7卷 12-15页

作者： Yuping WAN Xiaosheng WU Fangfang JIA Jing LI Zhaoqin WANG Yuejun FENG Meihong DU fangyang he Beijing Kwinbon Biotechnology Co. Ltd.Beijing 102206China Beijing Egineering Research Centre of Food Safety Immunodetection Beijing 102206China Beijing Center for Physical&Chemical Analysis Beijing 100089China

An immunomagnetic bead kit to separate and enrich salbutamol was prepared through the reaction between salbutamol and 4-aminobenzoic *** kit had a catch of 20 ng/ml to salbutamol in samples,and showed no cross reaction with competitive drugs with structure and function similar to salbutamol: clenbuterol,ractopamine,phenylethanolamine A, bromchlorbuterol, brombuterol, terbutaline, hydroxymethyl salbutamol,cimaterol,tulobuterol,mapenterol,cimbuterol,clenpenterol,zilpaterol,penbutolol,clenproperol,mabuterol and clorprenaline.

关键词： Salbutamol Monoclonal antibody Immunomagnetic bead kit to separate and enrich salbutamol

Preparation of a Salmonella LAMP Detection Kit

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Agricultural Biotechnology 2019年第6期8卷 97-99,103页

作者： Yuping WAN Xiaosheng WU Yu ZHANG Meihong DU Zhaoqin WANG Caiwei FENG Zhengmiao ZHAO fangyang he Beijing Kwinbon Biotechnology Co. Ltd.Beijing 102206China Beijing Engineering Technology Research Center of Food Safety Immune Rapid Detection Beijing 102206China Beijing center for physical&chemical analysis Beijing 100089China

[Objectives] This study was conducted to establish a method for rapid detection of Salmonella at the grassroots level, and a corresponding product was developed. [Methods] By analyzing the invA gene of Salmonella, two pairs of efficient, rapid and specific amplification primers were designed and screened for the six conserved domains of the gene, and the LAMP detection kit for Salmonella based on this method was produced. [Results] The method is simple, sensitive, rapid and specific. The minimum detection concentration is 10~2-10~3 CFU/ml. [Conclusions] It can be used in the self-inspection of food production enterprises and the on-site sampling inspection and rapid detection of large quantities of samples by grassroots food safety inspection and supervision departments.

关键词： LAMP Salmonella Rapid detection

饲料中硝基呋喃类药物免疫胶体金检测试纸条的研制

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饲料工业 2012年第23期33卷 35-38页

作者：何方洋冯才伟孙震赵正苗何丽霞北京勤邦生物技术有限公司北京102206

试验研究建立了一种基于免疫胶体金技术的硝基呋喃类药物快速检测方法。该方法对饲料中呋喃唑酮、呋喃它酮、呋喃西林、呋喃妥因的最低检测限分别达到5、3、2、2μg/kg,对饲料样本检测的假阳性率为2%,假阴性率为0,可以应用于饲料样本中... 详细信息

试验研究建立了一种基于免疫胶体金技术的硝基呋喃类药物快速检测方法。该方法对饲料中呋喃唑酮、呋喃它酮、呋喃西林、呋喃妥因的最低检测限分别达到5、3、2、2μg/kg,对饲料样本检测的假阳性率为2%,假阴性率为0,可以应用于饲料样本中硝基呋喃类药物的快速检测和现场筛查。

关键词：呋喃唑酮呋喃它酮呋喃西林呋喃妥因检测胶体金试纸条

三唑磷单克隆抗体制备以及酶联免疫试剂盒的研究

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中国酿造 2016年第6期35卷 165-169页

作者：冯才伟何方洋黄健贾芳芳李行北京勤邦生物技术有限公司北京102206 北京市食品安全免疫快速检测工程技术研究中心北京102206 北京市优质农产品产销服务站北京100101

通过三唑磷与乙酰氯反应,得到三唑磷半抗原,通过免疫动物得到抗三唑磷单克隆抗体,将其应用于能够检测蔬菜中三唑磷残留量的酶联免疫吸附测定（ELISA）试剂盒,结果表明：该试剂盒对结球甘蓝中三唑磷的检测限为49.6 μg/kg,IC50为6.9 μg... 详细信息

通过三唑磷与乙酰氯反应,得到三唑磷半抗原,通过免疫动物得到抗三唑磷单克隆抗体,将其应用于能够检测蔬菜中三唑磷残留量的酶联免疫吸附测定（ELISA）试剂盒,结果表明：该试剂盒对结球甘蓝中三唑磷的检测限为49.6 μg/kg,IC50为6.9 μg/L,加样回收率为70.0％～93.5％,三唑磷标准曲线线性范围为1～81 μ,g/L,批内、批间的相对标准偏差（RSD）＜ 10％,三唑磷单克隆抗体与毒死蜱、对硫磷的交叉反应率分别为2.3％,3.8％.4℃条件该试剂盒下能够保存12个月,稳定性较好.

关键词：三唑磷单克隆抗体 ELISA试剂盒

酶联免疫法与液相色谱-串联质谱法检测猪肉中沙丁胺醇

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中国酿造 2015年第8期34卷 132-135页

作者：何方洋于信念王坤王兆芹北京勤邦生物技术有限公司北京102206 北京市食品安全免疫快速检测工程技术研究中心北京102206 金元证券股份有限公司广东深圳518048

运用酶联免疫法（ELISA）与液相色谱-串联质谱法（LC-MS/MS）对猪肉中沙丁胺醇残留进行测定,并比较了二者在灵敏度、准确性和重现性等方面的差异。结果显示,ELISA法和LC-MS/MS法在猪肉样品中的检测限可达到0.5μg/kg和0.25μg/kg,分别... 详细信息

运用酶联免疫法（ELISA）与液相色谱-串联质谱法（LC-MS/MS）对猪肉中沙丁胺醇残留进行测定,并比较了二者在灵敏度、准确性和重现性等方面的差异。结果显示,ELISA法和LC-MS/MS法在猪肉样品中的检测限可达到0.5μg/kg和0.25μg/kg,分别向猪肉样品中添加3个质量浓度水平（1.0μg/kg、2.0μg/kg、4.0μg/kg）的沙丁胺醇时,回收率分别为83.7%～90.9%和86.6%～93.5%,变异系数（CV）分别为5.4%～10.3%和6.0%～8.3%。用酶联免疫法实际样品进行检测,筛选出2个阳性样品,经液相色谱-串联质谱法确证亦为阳性,测定结果一致。研究表明,酶联免疫法重复性较好、准确度较高,适用于进行猪肉样品中沙丁胺醇的快速筛选,液相色谱-串联质谱法适用于阳性样品的确证和精确定量。

关键词：猪肉沙丁胺醇检测酶联免疫法液相色谱-串联质谱法