Colloidal gold test strip used for detecting wheat leaf rust
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The technology of colloidal gold test paper and wheat leaf rust is applied in the field of wheat leaf rust prevention and control, which can solve the problems of time-consuming, laborious, complicated steps and the like, and achieve the effects of high titer, good specificity and strong affinity
Active Publication Date: 2015-10-28
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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[0005] However, the method is cumbersome and takes one day from sampling to obtaining results, which is time-consuming and laborious, and needs to be carried out in the laboratory
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Embodiment 1
[0041] Embodiment 1, the acquisition of wheat leaf rust monoclonal antibody
[0042] 1. Preparation of hybridoma cells
[0043] The experimental animals were BALB / c mice, provided by the Chinese Academy of Military Medical Sciences.
[0044] 1. Antigen preparation
[0045] (1) Use software to analyze the gene and amino acid sequence of wheat leaf rust protein to determine a certain number of amino acid sequences with antigenic sites, and synthesize them to obtain synthetic polypeptide products, and screen specific polypeptides. The sequence is:
[0046] Seq ID No. 1: CSSSKYQDSGREKQRTNSDSNDEIGA.
[0047] (2) The synthesized polypeptide product is used to couple protein BSA and protein OVA respectively, and use it as an immunogen and a coating source, wherein the immunogen is used to immunize mice, and the coating source is used for coating detection.
[0048] 2. Preparation of hybridoma cells
[0049] (1) Select mice with good serum detection of wheat leaf rust and perform...
Embodiment 2
[0075] Embodiment 2, be used to detect the preparation of the colloidal gold test strip of wheat leaf rust
[0076] According to the preparation method and process of colloidal gold test strips, corresponding auxiliary materials are prepared, which mainly include: NC film, PS bottom plate, gold standard pad, sample pad, absorbent paper (the above auxiliary materials are all in accordance with the existing standards of Beijing Qinbang Biotechnology Co., Ltd. Procurement and configuration of small molecule detection process).
[0077] (1) Preparation of gold particles
[0078] 1. Reagents:
[0079] 2% HAuCL4 (sigma) solution; the dilution method is double distilled water (the resistivity is about 18.2).
[0080] 3% trisodium citrate solution (sigma), which needs to be freshly prepared, and then filtered through a 0.35um membrane filter.
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Abstract
The invention provides a colloidal gold test strip used for detecting wheat leaf rust. The colloidal gold test strip comprises a sample pad, a colloidal-gold pad, an NC membrane, absorbent paper and a PS base plate, wherein the sample pad, the colloidal-gold pad, the NC membrane and the absorbent paper are successively arranged on the PS base plate along the chromatographic direction of a sample, the colloidal-gold pad is coated by a colloidal gold-labeled monoclonal antibody against Puccinia triticina, the NC membrane successively comprises a detection line and a quality control line along the chromatographic direction of the sample, the detection line is coated with Puccinia triticina antigen protein that the monoclonal antibody can specifically recognize, and the quality control line is coated with a goat anti-mouse secondary antibody. The monoclonal antibody against Puccinia triticina is secreted by a hybridoma cell with an accession number of CGMCC No. 10884. When the test strip is used for detection, the process from sampling to obtainment of detection results only takes 15 min, operation is simple and convenient, the results are accurate, early warning of wheat leaf rust can be realized, and economic loss caused by diseases is reduced.
Description
technical field [0001] The invention relates to the field of prevention and treatment of wheat leaf rust, in particular to a colloidal gold test strip for detecting wheat leaf rust. Background technique [0002] Wheat leaf rust (PucciniareconditaRob.exDesm.f.sp.triticiErikss.Et Henn.) is a fungal disease caused by wheat leaf rust, which is an important disease in China and all wheat-producing countries in the world. Can cause more than 15% yield loss. [0003] For a long time, the prediction of wheat rust has been mainly based on regular and large-scale field investigations and the isolation, cultivation and identification of indoor pathogen races. Not only is it time-consuming and labor-intensive, but the accuracy and reliability of the results also depend to a large extent on the technical level and experience of the surveyors, making it difficult to meet the actual needs of rapid, high-throughput diagnostic testing. The difference in seedling symptoms between leaf rust ...
Claims
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