Rapid immunoassay test strip for bacterial fruit blotch II type strains of melons and application thereof
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A rapid immunization technology for fruit spot bacteria, applied in immunoassays, biological tests, measuring devices, etc., can solve the problems of differences in fungicide sensitivity and pathogenicity, achieve good stability, short detection time, and reduce field The effect of sample size
Inactive Publication Date: 2020-10-02
NANJING AGRICULTURAL UNIVERSITY
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Although different subgroups can induce plant disease, there are significant differences in their pathogenicity. The pathogenicity of subgroup II strains to watermelon hosts is stronger than that of other hosts, while subgroup I strains are more pathogenic to melon hosts. are moderately infested
Moreover, the susceptibility of different subgroup strains to fungicides varies
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Embodiment 1
[0027] Embodiment 1: Specificity test of rapid immunoassay test strips of melon bacterial fruit blotch type II strain
[0028] 1. Identification of the tested strains by gene molecular technology (PCR)
[0029] A total of 90 strains for testing were collected from crops with suspected symptoms all over the country, and 90 strains were selected with BX-L1 / BX-S-R2 general primers and G2-Fwd / G12-Fwd / G32-Rwd specific primer pairs Perform PCR amplification. PCR reaction system (25 μL): PCR reaction system (25 μl): 10×PCR buffer 2.5 μl, Mg 2+ 2 μl, 0.5 μl of d NTP, 0.3 μl of Taq enzyme, 0.3 μl of forward and reverse primers, 1 μl of glycerol bacterial liquid as template, 18.1 μl of sterile three-distilled water, and the standard Xinjiang fruit spot strain as a positive control.
[0030] PCR reaction conditions: 30 cycles of pre-denaturation at 95°C for 5 min, denaturation at 95°C for 30 s, annealing at 55°C for 40 s, extension at 72°C for 30 s in a PCR machine, incubation at 72°C ...
Embodiment 2
[0040]Example 2: Test of field samples by rapid immunoassay test strips of melon bacterial fruit spot bacillus type II strains
[0041] 1. Field sampling and processing
[0042] Collect the suspected symptoms of leaves of melon, watermelon and other crops in the field, tear out a sample of about 5mm×5mm (at the junction of disease and health) and put it in a single-sided frosted grinding bag, fold the frosted side of the grinding bag in half along the middle, and place the sample on the folded surface Grind thoroughly in the medium, add 2mL of the sample filling solution, mix well and be tested.
[0043] Use a plastic dropper to draw 100-200 μL of the test solution, drop it into the sample pad of the test strip, react for 5-10 minutes, and judge the result according to the following rules.
[0044] (1) The quality control line (C) develops color, and the test line (T) has a red band, which is positive
[0045] (2) The C line develops color, and the T line does not have a red...
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Abstract
The invention discloses a rapid immunoassay test strip for bacterial fruit blotch II type strains of melons and application thereof. The test strip comprises a bottom plate, wherein a water absorptionpad, 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 atthe 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-cucurbit bacterial fruit blotch germ polyclonal antibody 1152, and the quality control line iscoated with goat anti-mouse IgG. The gold label pad is sprayed with a nanogold-labeled monoclonal antibody for resisting bacterial fruit blotch II type strains of melons. The test strip is high in specificity, capable of distinguishing bacterial fruit blotch type I strains and bacterial fruit blotch type II strains of melons, free of cross reaction with bacterial angular leaf blotch and the like,capable of achieving the sensitivity of 1*10<5> cfu / mL and capable of being used for rapid screening of the bacterial fruit blotch type II strains of the melons in the field.
Description
technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a rapid immunological detection test strip of melon bacterial fruit spot pathogen type II strain and an application thereof. Background technique [0002] Bacterial fruit blotch (BFB) of melons is a typical seed-borne quarantine disease caused by bacterial fruit blotch of melons (Acidovorax citrulli, A.citrulli). After being introduced into my country from abroad or outside the region, the disease has occurred in almost all melon planting areas in my country, posing a serious threat to the production of cucurbit crops such as muskmelon and watermelon in my country, and causing huge economic losses to local farmers. [0003] Bacterial leaf spot of melons can be divided into subgroup I and subgroup II, which have rich genetic diversity. Although different subgroups can induce plant disease, there are significant differences in their pathogenicity. The pathogenicity ...
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