A kind of in situ ELISA quantitative detection method of residual bt protein in soil

A quantitative detection method, bt protein technology, applied in the field of protein detection

Active Publication Date: 2016-09-14
NANJING INST OF ENVIRONMENTAL SCI MINIST OF ECOLOGY & ENVIRONMENT OF THE PEOPLES REPUBLIC OF CHINA
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Problems solved by technology

[0004] Purpose of the invention: Aiming at the disadvantages of the existing soil Bt protein detection methods, the present invention has developed a new method for in situ ELISA quantitative detection of bt protein directly on the surface of soil particles, which can be directly and accurately detected without extraction. Detect the difficult-to-extract bt protein adsorbed on the surface of soil particles, which truly reflects the residual situation of Bt protein in the soil

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  • A kind of in situ ELISA quantitative detection method of residual bt protein in soil

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Embodiment 1

[0036] The soil samples used in this program were taken from the cotton fields planted with Bt cotton and conventional cotton in Nanjing area for one year, two years and five years respectively, and the sampling site was Liuhe, Nanjing.

[0037] During the flowering stage of cotton, soil samples were randomly collected from Bt cotton fields and conventional cotton fields, and the 5-point sampling method was adopted, and 5 sampling points were set for each cotton field. Each sampling point is 30-50m apart, and 5 cotton plants are randomly selected around each sampling point. Around the rhizosphere of each cotton plant (within 5cm from the main root, 5-20cm from the surface), a total of about 500g of soil is taken and packed into a sealed container. bag, as a test sample. The collected samples were brought back to the laboratory and stored in a -20°C refrigerator for testing.

[0038] Take 1-2g of soil and proceed as follows:

[0039] 1. Remove the animal and plant residues in...

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Abstract

The invention discloses an in-situ enzyme linked immunosorbent assay quantitative determination method of a Bt protein residual in soil. The method comprises 1, removing animal and plant residues in a soil sample to be detected, carrying out full grinding, and screening the soil sample by a sieve of 50-60 meshes so that soil particle sizes are 0.2mm or less, 2, carrying out sterilization on the ground soil sample under conditions of high pressure, dampness and heat for 15-20min, 3, weighing 1-2g of the soil sample subjected to sterilization, putting the soil sample into a centrifuge tube with a volume of 10ml, and carrying out washing to remove impurities, 4, enclosing the surface of the soil sample without impurities, and 5, carrying out antibody fixation detection on the enclosed soil sample. The method provides a novel approach for monitoring soil Bt protein residue and Bt protein residue-caused influence on a soil ecosystem, can be used for detecting soil Bt protein and can also be used for detecting various soil residual protein contents after replacement with different antibodies.

Description

technical field [0001] The invention relates to the technical field of protein detection, in particular to an in-situ ELISA quantitative detection method for residual Bt protein in soil. Background technique [0002] In recent years, with the continuous expansion of the planting area of ​​Bt crops, their potential risks to the ecological environment have increasingly become a hot topic of research by experts and scholars at home and abroad. After planting, transgenic Bt crops will release Bt toxins to the soil through root exudates, pollen, and plant residues throughout their growth period, and these secreted Bt toxins will be adsorbed by surface active particles such as clay minerals and humus in the soil , and the bound toxin still has strong insecticidal activity and can persist in the soil environment for a long time. Soil is an important place for material cycle and energy conversion in the ecosystem, and the animal and microbial populations in the soil play a very imp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535
CPCG01N33/535
Inventor 方志翔刘标沈文静李衍亮
Owner NANJING INST OF ENVIRONMENTAL SCI MINIST OF ECOLOGY & ENVIRONMENT OF THE PEOPLES REPUBLIC OF CHINA
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