Colloidal gold test strip used for detecting wheat leaf rust
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
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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.
[0081] Instrument: electromagnetic heating stirring jacket thermostat + whirling flow round flask + snake-shaped condenser tube + stirring ...
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