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Light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis

A photo-excited chemiluminescence and immunodetection technology, which is applied in the field of agricultural biology, can solve the problems of no report on the detection of Microsporidia Bombyx by AlphaLISA, and achieve the effects of high uniformity, high sensitivity and accuracy, and less background interference.

Active Publication Date: 2015-08-26
SOUTHWEST UNIVERSITY +1
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Problems solved by technology

However, there are no reports at home and abroad on the detection of AlphaLISA for N. silkworm

Method used

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  • Light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis
  • Light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis
  • Light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis

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Embodiment Construction

[0031] Below in conjunction with accompanying drawing and specific embodiment, the present invention is described in further detail, if there is no special specification in the raw material used in the embodiment, be common commercially available; Adopted biological experiment is if no special specification, is conventional method.

[0032] One, a kind of photoactivated chemiluminescence immunoassay method for silkworm finished egg microsporidiosis, comprises the following steps:

[0033] 1. Establishment of hybridoma cell lines 1G5 and 2D9 of mouse anti-Bombyx mori monoclonal antibody to Microsporidium sporophytum protein;

[0034] 2. Preparation and purification of the ascites of the monoclonal antibody 1G5 and 2D9 monoclonal antibodies to No. spp. mori;

[0035] 3. Detection of AlphaLISA for Microsporidium silkworm;

[0036] 4. Evaluation of AlphaLISA detection method.

[0037] 2. The specific steps include:

[0038] 1. Establishment of a mouse hybridoma cell line with a...

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Abstract

The invention provides a light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis. The light-stimulating chemiluminiscence immunological detection method comprises preparation of silkworm microsporidia sporoderm protein antigens, establishment of anti-rat silkworm microsporidia sporoderm protein monoclonal antibody hybridoma cell strains, ascites preparation and purification of silkworm microsporidia sporoderm protein monoclonal antibodies, and establishment of a silkworm microsporidia Alpha LISA detection method. When the method is used for detecting, monomer oxygen cannot be dispersed to receptor microspheres when biomolecules do not have specific mutual effect, and no signals are generated, so that the relatively high sensitivity and accuracy are realized, and a detection result is relatively accurate; the method has relatively high uniformity, less background interference and low background, and has a wide dynamic detection range; and when a kit is used for detecting, the demanded quantity of samples is extremely few and the samples do not need to be washed in a detection process, so that a detection flow is simplified and the market prospect is good.

Description

technical field [0001] The invention belongs to the technical field of agricultural biology, and in particular relates to a photochemiluminescent immunodetection method for the microsporidiosis of silkworm finished eggs. Background technique [0002] The microsporidium disease caused by Nosema bombycis (Nb) infecting silkworm is a destructive infectious disease of silkworm, and it has been identified as the only quarantine disease in sericulture areas in the world. In recent years, scholars at home and abroad have carried out a lot of research work on the detection of silkworm microparticle disease, and made great progress. However, due to the influence of cost, actual sensitivity, and operational complexity, there has not been a technology that can replace the current optical method. Microscopic detection method. AlphaLISA technology is a new type of homogeneous detection technology based on bead-based chemiluminescence. The detection principle of AlphaLISA is similar to t...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/569G01N33/577G01N33/543
Inventor 吴胜昔潘国庆周泽扬李春峰吴海晶李曾蔡家利
Owner SOUTHWEST UNIVERSITY
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