Chinese Academy of Agricultural Sciences

Oilcrops Research Institute of Chinese Acadamy of Agriculture Sciences

Chinese Academy of Sciences

Wuhan Norrida Biotechnological Co., Ltd

Zhaowei Zhang

Peiwu Li

Qi Zhang

Xiaoxia Ding

Wen Zhang

Ting He

Jun Jiang

Xiaoqian Tang

Du Wang

Min Li

Xiao Chen

Li Yu

Xiaofeng Hu

Xin Li

Wenqin Wu

Ling Cheng

Xiupin Wang

Yanru Wang

Ran Li

A time-resolved fluorescence immunochromatography kit for the simultaneous detection of a mixed pollutant of aflatoxin and carbaryl, a preparation method therefor, and an application thereof. The kit comprises a time-resolved fluorescence immunochromatography test strip and a sample reaction vial containing a europium-labeled anti-aflatoxin monoclonal antibody and a europium-labeled anti-carbaryl monoclonal antibody lyophilized product, detection lines of the time-resolved fluorescence immunochromatography test strip being coated with an aflatoxin-bovine serum albumin conjugate and a carbaryl-ovalbumin conjugate respectively, and the anti-carbaryl monoclonal antibody is produced by secretion of a hybridoma cell line Jnw1D2 with a preservation number of CCTCC NO. C201654. The kit can be used for the simultaneous detection of aflatoxin and carbaryl content in a sample, and features simple and quick operations and a high sensitivity.

Time-resolved fluorescence immunochromatography kit for simultaneous detection of mixed pollutant of aflatoxin and carbaryl, preparation method therefor, and application thereof

A time-resolved fluorescence immunochromatographic kit for simultaneous detection of mixed contamination of five mycotoxins such as aflatoxin B1 and an application thereof are disclosed. The kit comprises a time-resolved fluorescent immunochromatographic test strip and sample reaction vials each containing a europium-labeled monoclonal antibody lyophilized product; wherein the fluorescent test strip comprises a PVC substrate, and a surface of the PVC substrate is adhered with a water absorbing pad (), a detection pad () and a sample pad () from top to bottom, adjacent pads being connected and overlapping at connections. The detection pad () adopts a nitrocellulose membrane as the base thereof and is arranged with a lateral quality control line () and five detection lines () from top to bottom each covered by a bovine serum albumin conjugate of each toxin. The fumonisin B1 monoclonal antibody is generated by the hybridoma cell strain Fm7A11 having China Center for Type Culture Collection (CCTCC) accession number C201636. The kit is applicable to simultaneous detection of mixed contamination of aflatoxin B1, fumonisin B1, ochratoxin A, zearalenone and sterigmatocystin.

Time-resolved fluorescence immunochromatographic kit for simultaneous detection of mixed contamination of five mycotoxins such as aflatoxin B1 and application thereof

The present disclosure belongs to the technical field of biosensors and particularly provides a construction method for a photocathode indirect competition sensor and an evaluation method. The construction method includes: using Z-type BiO/CuBiOas a sensing platform; calculating a photoinduced electron Z-type transfer path and an energy band structure of BiOand CuBiOusing a density functional theory (DFT); and constructing a BiO/CuBiO-based biosensor. A photoelectrochemical (PEC) photocathode biosensor based on a BiO/CuBiOheterojunction prepared through the solution has good repeatability, reproducibility, stability, and specificity for detecting a target. The PEC biosensor constructed in the solution of the present disclosure has a broad application prospect in the fields of healthcare, environment, and food.

Construction method for photocathode indirect competition sensor and evaluation method

The present disclosure discloses an intelligent point-of-care testing device for mycotoxins based on a quantum dots test strip. A first light source and a second light source are used for exciting a test strip which is placed in a shell and coated with a mycotoxin antigen to obtain emitted light, and a light filtering component is used for filtering out impurity light. A clamping mechanism is used for clamping and fixing an image capturing component. The image capturing component is used for capturing, an image of mycotoxins based on a quantum dots test strip. A processor is used for analyzing and calculating image information to obtain a mycotoxin content result. The testing time is shortened, and the portability and practicability of mycotoxin testing are improved. Quantitative instant testing of single or multi-toxin mixed pollution is achieved, and the technical problem of fluorescence quantification is solved.

Intelligent instant quantitative testing device for mycotoxin quantum dot test strip

An immunoadsorbent for purifying fumonisin B, aflatoxin B, ochratoxin A, zearalenone and sterigmatocystin; a complex affinity column and its preparation method; and a method for detecting the mycotoxins using the same are provided. The immunoadsorbent comprises a solid-phase support, and an anti-fumonisin Bmonoclonal antibody, an anti-aflatoxin Bmonoclonal antibody, an anti-ochratoxin A monoclonal antibody, an anti-zearalenone monoclonal antibody and an anti-sterigmatocystin monoclonal antibody which are coupled to the solid-phase support, wherein the anti-fumonisin Bmonoclonal antibody is secreted by a hybridoma cell strain Fm7A11. The complex affinity column can be used for purification and detection of a fumonisin Bsample, an aflatoxin Bsample, an ochratoxin A sample, a zearalenone sample and a sterigmatocystin sample at the same time.

Immunoadsorbent for purifying five kinds of mycotoxins including fumonisin Band aflatoxin B, and complex affinity column

An aflatoxin nanobody immunoabsorbent and immunoaffinity column and preparation method and use thereof. The immunoabsorbent comprises a solid phase carrier and aflatoxin B1 nanobody 2014AFB-G15 coupled with the solid phase carrier. The 50% inhibiting concentration ICof aflatoxin B1 nanobody 2014AFB-G15 to aflatoxin B1 is 0.66 ng/mL, and the cross-reactivity of aflatoxin B1 nanobody 2014AFB-G15 to aflatoxins B2, G1, G2, and M1 are respectively 22.6%, 10.95%, 32.1% and 26%. The amino acid sequence of aflatoxin B1 nanobody 2014AFB-G15 is as depicted by SEQ ID NO: 7, and the coding gene sequence thereof is as depicted by SEQ ID NO: 8. The aflatoxin nanobody immunoaffinity column can be used for purification and concentration of sample extract prior to computer testing, and the immunoaffinity column can be reused repeatedly.

Aflatoxin nanobody immunoabsorbent and immunoaffinity column and preparation method and use thereof

An aflatoxin M1 nanobody, an immunosorbent and an immunoaffinity column. The aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, is encoded by the nucleic acid sequence of SEQ ID NO:8, has a 50% inhibiting concentration ICto aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxins B1, B2, G1, and G2 of 9.43%, 5.93%, 4.87% and 6.17%, respectively. The immunosorbent includes a solid phase carrier and aflatoxin M1 nanobody 2014AFM-G2 coupled with the solid phase carrier. The immunoaffinity column is loaded with the aflatoxin M1 nanobody immunosorbent. It can be used for purifying and concentrating an extracting solution of a sample before loading to a machine for detection and the immunoaffinity column can be used repeatedly for many times.

Immunosorbent and immunoaffinity column for aflatoxin M1 nanobody and preparation method thereof

A hybridoma cell line ST03 having China Center for Type Culture Collection (CCTCC) accession number C2013187, a monoclonal antibody against aflatoxin biosynthetic precursor ST produced by the hybridoma cell line ST03, and the use of the monoclonal antibody. The hybridoma cell line ST03 can be used for preparing a high-titer monoclonal antibody against aflatoxin biosynthetic precursor ST, and the titer of mouse ascites antibody against aflatoxin biosynthetic precursor ST determined by enzyme linked immunosorbent assay (ELISA) can reach 6.4&#xd7;10. The monoclonal antibody against aflatoxin biosynthetic precursor ST has high sensitivity, has 50% inhibiting concentration ICto aflatoxin biosynthetic precursor ST of 0.36 ng/mL, has no cross reaction with all of aflatoxin B1, aflatoxin B2, aflatoxin G1, and aflatoxin G2, and can be used for the content determination of aflatoxin biosynthetic precursor ST.

Hybridoma cell line ST03, monoclonal antibody against aflatoxin biosynthetic precursor sterigmatocystin and use thereof

Aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, and is encoded by the gene sequence of SEQ ID NO:8. The aflatoxin M1 nanobody 2014AFM-G2 obtained via screening has the properties of tolerance to organic reagents, tolerance to high temperature, tolerance to acids and bases and the like, and good stability. The aflatoxin M1 nanobody 2014AFM-G2 has 50% inhibiting concentration ICto aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxin B1, B2, G1, G2 are 9.43%, 5.93%, 4.87% and 6.17%, respectively.

Aflatoxin M1 nanobody 2014AFM-G2

Hybridoma cell line 10G4 and monoclonal antibody against total aflatoxins produced by the hybridoma cell line 10G4. The hybridoma cell line 10G4 is used to produce the monoclonal antibody that binds specifically total aflatoxin B1, B2, G1 and G2. The titer of the mouse ascites antibody produced by the 10G4 treated mouse is determined through non-competitive enzyme-linked immunosorbent assay and the titer can reach up to 5.12&#xd7;10. The monoclonal antibody against total aflatoxin B1, B2, G1 and G2 are used for better identification of aflatoxin B1, B2, G1 and G2 with good consistency. The 50% inhibitory concentrations (IC) of the antibody against aflatoxin B1, B2, G1 and G2 are 2.09 ng/mL, 2.23 ng/mL, 2.19 ng/mL and 3.21 ng/mL respectively. The range of cross reaction rate for aflatoxin B1, B2, G1 and G2 is about 65.2%-100%. The antibody is used for quantitative measurement of total aflatoxin B1, B2, G1 and G2.

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Hubei, China

Zhaowei Zhang