Anti-Idiotypic Nanobody-Phage Based Real-Time Immuno-PCR for Detection of Hepatocarcinogen Aflatoxin in Grains and Feedstuffs
Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-produ...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2014-11, Vol.86 (21), p.10841-10846 |
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| creator | Lei, Jiawen Li, Peiwu Zhang, Qi Wang, Yanru Zhang, Zhaowei Ding, Xiaoxia Zang, Wen |
| description | Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R 2 = 0.991) were obtained. In this context, this study provides a novel opportunity to analyze aflatoxins in agri-products. |
| doi_str_mv | 10.1021/ac5029424 |
| format | Article |
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Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R 2 = 0.991) were obtained. 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Chem</addtitle><description>Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R 2 = 0.991) were obtained. In this context, this study provides a novel opportunity to analyze aflatoxins in agri-products.</description><subject>Aflatoxins</subject><subject>Aflatoxins - analysis</subject><subject>Aflatoxins - toxicity</subject><subject>Animal Feed - analysis</subject><subject>Antibodies, Anti-Idiotypic - immunology</subject><subject>Arachis hypogaea</subject><subject>Assaying</subject><subject>Base Sequence</subject><subject>Carcinogens</subject><subject>Carcinogens - analysis</subject><subject>Carcinogens - toxicity</subject><subject>Corn</subject><subject>DNA Primers</subject><subject>Edible Grain - chemistry</subject><subject>Feeds</subject><subject>Liver Neoplasms - chemically induced</subject><subject>Nanoparticles</subject><subject>Nanostructure</subject><subject>Oryza sativa</subject><subject>Peanuts</subject><subject>Phage</subject><subject>Polymerase chain reaction</subject><subject>Real time</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Single-Domain Antibodies - immunology</subject><subject>Toxins</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0UFrFDEYBuAgit2uHvwDEhChHqLJl0kyc1y3tl0oWko9D5lMUlNmkm0yA91D_3tTthbRixAIhIf3-8KL0DtGPzMK7Is2gkJTQfUCLZgASmRdw0u0oJRyAorSA3SY8w2ljFEmX6MDEKA4F7BA96swebLpfZx2W2_wdx1iF_sdufilry3-qrPt8aXVA7nyo8WbcZxDJBfrS-xiwsd2smbyMeDo8Jnd6ikanYwP8doGvHJDebjzAZdzmrQPGevQ4xNr-zzNzuU36JXTQ7Zvn-4l-nny7Wp9Rs5_nG7Wq3OiKyUmIphhuumUs1ApqYXsNHAAChKsVI3gqna9BKVUMbLunDFSNh3jTvG6ow1foqN97jbF29nmqR19NnYYdLBxzi2TDXDJaiX-g4JqFKhKFfrhL3oT5xTKRx5V2VDWpYAl-rRXJsWck3XtNvlRp13LaPtYX_tcX7HvnxLnbrT9s_zdVwEf90Cb_Me0f4IeAAj6nkw</recordid><startdate>20141104</startdate><enddate>20141104</enddate><creator>Lei, Jiawen</creator><creator>Li, Peiwu</creator><creator>Zhang, Qi</creator><creator>Wang, Yanru</creator><creator>Zhang, Zhaowei</creator><creator>Ding, Xiaoxia</creator><creator>Zang, Wen</creator><general>American Chemical Society</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7TV</scope></search><sort><creationdate>20141104</creationdate><title>Anti-Idiotypic Nanobody-Phage Based Real-Time Immuno-PCR for Detection of Hepatocarcinogen Aflatoxin in Grains and Feedstuffs</title><author>Lei, Jiawen ; Li, Peiwu ; Zhang, Qi ; Wang, Yanru ; Zhang, Zhaowei ; Ding, Xiaoxia ; Zang, Wen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a475t-51c1a9b7fe2476a56ba23220262e6795378fd62777b7f68bfcc669b13f738b093</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Aflatoxins</topic><topic>Aflatoxins - analysis</topic><topic>Aflatoxins - toxicity</topic><topic>Animal Feed - analysis</topic><topic>Antibodies, Anti-Idiotypic - immunology</topic><topic>Arachis hypogaea</topic><topic>Assaying</topic><topic>Base Sequence</topic><topic>Carcinogens</topic><topic>Carcinogens - analysis</topic><topic>Carcinogens - toxicity</topic><topic>Corn</topic><topic>DNA Primers</topic><topic>Edible Grain - chemistry</topic><topic>Feeds</topic><topic>Liver Neoplasms - chemically induced</topic><topic>Nanoparticles</topic><topic>Nanostructure</topic><topic>Oryza sativa</topic><topic>Peanuts</topic><topic>Phage</topic><topic>Polymerase chain reaction</topic><topic>Real time</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Single-Domain Antibodies - immunology</topic><topic>Toxins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lei, Jiawen</creatorcontrib><creatorcontrib>Li, Peiwu</creatorcontrib><creatorcontrib>Zhang, Qi</creatorcontrib><creatorcontrib>Wang, Yanru</creatorcontrib><creatorcontrib>Zhang, Zhaowei</creatorcontrib><creatorcontrib>Ding, Xiaoxia</creatorcontrib><creatorcontrib>Zang, Wen</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Pollution Abstracts</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lei, Jiawen</au><au>Li, Peiwu</au><au>Zhang, Qi</au><au>Wang, Yanru</au><au>Zhang, Zhaowei</au><au>Ding, Xiaoxia</au><au>Zang, Wen</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Anti-Idiotypic Nanobody-Phage Based Real-Time Immuno-PCR for Detection of Hepatocarcinogen Aflatoxin in Grains and Feedstuffs</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2014-11-04</date><risdate>2014</risdate><volume>86</volume><issue>21</issue><spage>10841</spage><epage>10846</epage><pages>10841-10846</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R 2 = 0.991) were obtained. In this context, this study provides a novel opportunity to analyze aflatoxins in agri-products.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>25273352</pmid><doi>10.1021/ac5029424</doi><tpages>6</tpages></addata></record> |
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| subjects | Aflatoxins Aflatoxins - analysis Aflatoxins - toxicity Animal Feed - analysis Antibodies, Anti-Idiotypic - immunology Arachis hypogaea Assaying Base Sequence Carcinogens Carcinogens - analysis Carcinogens - toxicity Corn DNA Primers Edible Grain - chemistry Feeds Liver Neoplasms - chemically induced Nanoparticles Nanostructure Oryza sativa Peanuts Phage Polymerase chain reaction Real time Real-Time Polymerase Chain Reaction - methods Single-Domain Antibodies - immunology Toxins |
| title | Anti-Idiotypic Nanobody-Phage Based Real-Time Immuno-PCR for Detection of Hepatocarcinogen Aflatoxin in Grains and Feedstuffs |
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