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Rapid immunoassay test strip for phytophthora infestans and application thereof

A technology for rapid immunization of E. amylovora, applied in immunoassays, biological tests, measuring devices, etc., can solve problems that cannot fully meet the needs of rapid inspection and quarantine, improve the efficiency of rapid screening, reduce the amount of field sampling, The effect of short detection times

Inactive Publication Date: 2020-10-02
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Gene molecular level detection technology has the advantages of accuracy and sensitivity, but requires relatively strict experimental conditions, measures and operations, and cannot fully meet the existing needs of rapid inspection and quarantine

Method used

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  • Rapid immunoassay test strip for phytophthora infestans and application thereof

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Experimental program
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Embodiment 1

[0022] Embodiment 1: Sensitivity test of E. amylovora rapid immunity test strip

[0023] Take E. amylovora, streak on LB plate medium, and place it in a constant temperature incubator at 28°C for cultivation. After 1-3 days of culture, after colonies grow, pick a single colony and inoculate it in NA medium, and place it on a shaker (28°C, 200rpm) for cultivation. When the bacterium grows to an OD of 0.3, take the cultured bacterium solution and centrifuge at 4°C and 6000rpm for 5min, wash twice with test strip chromatography solution and suspend to prepare 10 8 cfu / mL bacterial suspension, ready for use.

[0024] Dilute the above test solution gradient to: 10 8 、10 7 、10 6 、10 5 、10 4 For the CFU / mL gradient bacterial solution to be tested, take 150 μl of the gradient bacterial solution to be tested and drop it into the sample pad of the rapid immunoassay test strip for E. amylovora. After 5-10 minutes of chromatography, judge the pear according to the color of the test ...

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Abstract

The invention discloses a rapid immunoassay test strip for phytophthora infestans and application thereof. The test strip comprises a bottom plate, wherein a water absorption pad, a detection pad, a gold label pad and a sample pad are sequentially adhered to one surface of the bottom plate from top to bottom; every two adjacent pads are connected in an overlapped mode at the joint, a nitrocellulose membrane serves as a base pad of the detection pad, a quality control line and a detection line are transversely arranged on the nitrocellulose membrane from top to bottom, the detection line is located below the quality control line, the detection line is coated with an anti-pyricularia grisea polyclonal antibody 0550, and the quality control line is coated with goat anti-mouse IgG. The gold label pad is sprayed with a nanogold labeled anti-pyricularia oryzae monoclonal antibody. The specificity is high, and no cross reaction is caused to pythium aphanidermatum, pear rust water, pear rot, pear anthracnose and the like of Asian pears; the sensitivity of the test strip can reach 1*10<5> cfu / mL, and the test strip can be used for rapid screening of pythium aphanidermatum in fields.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a rapid immunological detection test strip for E. amylovora and its application. Background technique [0002] Fireblight, caused by Erwinia amylovora, is a devastating bacterial disease on pome fruit trees such as pears and apples. The disease first occurred in the United States, and then spread and spread through various channels. So far, the disease has spread to more than 40 countries in Central Europe, Southeast Europe, the Mediterranean Sea and Australia. According to the risk assessment results of relevant experts, most of the pear and apple production areas in my country are suitable for the growth of fire blight bacteria, and most of the local and imported pear and apple varieties planted in my country are highly susceptible varieties. Therefore, once the fire blight bacteria is introduced, in addition to the loss caused by direct damage, most of the inte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/58G01N33/558G01N33/52
CPCG01N33/577G01N33/56911G01N33/587G01N33/558G01N33/52G01N2469/10
Inventor 王利民田艳丽王岩李潇楠姜培郑秋菊胡白石
Owner NANJING AGRICULTURAL UNIVERSITY
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