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Immune colloidal gold test strip and detection method for karyotype polyhedrosis viruses of bombyx mori

A nuclear polyhedron and colloidal gold test paper technology is applied in the field of virus epidemic diagnosis to achieve the effect of ensuring specificity, strong pertinence, and strong practicability

Inactive Publication Date: 2015-03-25
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, this technology has not been reported in the detection of silkworm nuclear polyhedrosis virus (BmNPV), and because the hemolymph and tissue of the silkworm are convenient to collect, especially the hemolymph is not easy to coagulate, the hemolymph of the silkworm can be directly collected for colloid In the detection of gold, it can greatly reduce the operation steps and use costs, and is more convenient for silkworm farmers and front-line technicians to master and use. Therefore, the diagnostic colloidal gold test strip has a good application prospect

Method used

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  • Immune colloidal gold test strip and detection method for karyotype polyhedrosis viruses of bombyx mori
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  • Immune colloidal gold test strip and detection method for karyotype polyhedrosis viruses of bombyx mori

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Embodiment 1

[0037] 1. Material

[0038] Bombyx mori nuclear polyhedrosis virus, mouse anti-bombyx mori nuclear polyhedrosis virus monoclonal antibody (named BMlef4), rabbit anti-bombyx mori nuclear polyhedrosis virus polyclonal antibody were prepared and preserved by our laboratory. Trisodium citrate, chloroauric acid (HAuCl 4 4H 2 O) was purchased from Sigma Company; BSA was purchased from Promega Company; nitrocellulose membrane (NC) and glass fiber paper were purchased from Millipore Company; goat anti-mouse IgG was purchased from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.

[0039] 2. Preparation of BmNPV immune colloidal gold test strips

[0040] (1) First, the highly conserved early core gene Lef4 (GenBank: AY033655.1) of Bombyx mori nuclear polyhedrosis virus was selected for cloning, expression and purification to obtain the exogenous protein BmNPV-Lef4.

[0041] (2) Preparation of polyclonal antibody to BmNPV-Lef4 antigen:

[0042] Using the purified protein BmNPV-Lef...

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Abstract

The invention relates to an immune colloidal gold test strip and a detection method for karyotype polyhedrosis viruses of bombyx mori, particularly relates to an immune colloidal gold diagnosis test strip for detecting the karyotype polyhedrosis viruses of the bombyx mori by adopting an immunochromatography technology, and a preparation method of the immune colloidal gold diagnosis test strip, and belongs to the technical field of virus epidemic disease diagnosis. The immune colloidal gold test strip comprises a colloidal gold pad marked with an antigen BmNPV-Lef4 antibody and a coated nitrocellulose membrane, wherein an upper detection line of the nitrocellulose membrane is coated by rabbit-anti BmNPV-Lef4; and a quality control line at the lower part of the nitrocellulose membrane is coated with goat-anti-mouse IgG. The immune colloidal gold test strip is used for detecting the karyotype polyhedrosis viruses of the bombyx mori; compared with a traditional colloidal gold detection method, a double-antibody sandwich method is adopted when the colloidal gold test strip is prepared and a concentration proportion of two virus antibodies is regulated, so that the specificity, the sensitivity and the stability of a detection result can be effectively guaranteed; the detection sensitivity is high and the detection method is simple and convenient and is applied to the detection of the karyotype polyhedrosis viruses of the bombyx mori for the first time.

Description

technical field [0001] The present invention relates to an immune colloidal gold detection test strip for silkworm nuclear polyhedrosis virus and a detection method thereof, in particular to an immune colloidal gold diagnostic test strip for detecting silkworm nuclear polyhedrosis virus by an immune chromatography technique and a preparation method thereof, The invention belongs to the technical field of virus epidemic diagnosis. Background technique [0002] The silkworm is an important economic insect, and the silk it produces is the main source of silk raw materials, and plays an important role in human economic life and history and culture. However, the high incidence of silkworm disease directly affects the yield and quality of silkworm cocoons, restricting the healthy and sustainable development of sericulture. Silkworm diseases are divided into infectious silkworm diseases and non-infectious silkworm diseases, with infectious silkworm diseases being the most importan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/558G01N33/56983
Inventor 周阳施海峰高力刘海涛夏恒传刘晓勇姚勤
Owner JIANGSU UNIV
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