Primer for quickly and quantitatively detecting Wolbachia in tissues of mosquito and reagent box and method thereof

A technology for quantitative detection and detection of primers, applied in the biological field, can solve the problems of limited detection conditions, time-consuming, low specificity, etc., and achieve the effects of saving time and cost, improving work efficiency, and easy operation

Active Publication Date: 2012-03-21
GUANGZHOU WOLBAKI BIOTECH CO LTD
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation of antibodies is not only time-consuming but also time-consuming. Generally, it takes at least 2-3 months to prepare antibodies with low specificity (polyclonal antibodies), and it takes longer to prepare antibodies with high specificity (monoclonal antibodies).
At the same time, this detection condition is very restrictive, and multiple samples cannot be detected at the same time, so large-scale detection cannot be performed.
In addition, the biggest disadvantage of this method is that it cannot quantitatively analyze Wolbachia

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer for quickly and quantitatively detecting Wolbachia in tissues of mosquito and reagent box and method thereof
  • Primer for quickly and quantitatively detecting Wolbachia in tissues of mosquito and reagent box and method thereof
  • Primer for quickly and quantitatively detecting Wolbachia in tissues of mosquito and reagent box and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Quantitative determination of Wolbachia (Wolbachia) in midgut, salivary gland, ovary tissue of Aedes aegypti artificially transfected type B Wolbachia (Wolbachia)

[0024] 1. Acquisition of mosquito tissue samples containing Wolbachia: use CO 2 Anesthetize the mosquito to be tested (Aedes aegypti mosquito artificially transfected with Wolbachia type B), dissect and separate the mosquito tissue (midgut, salivary gland, ovary) to be tested with a needle under a dissecting microscope, and use STE buffer The isolated tissue was rinsed three times and then placed in 50 μL STE buffer. The STE buffer solution belongs to the prior art buffer solution, and its formula is 100mM NaCl, 10mM Tris-Cl, pH 8.0 and 1mM EDTA, pH 8.0.

[0025] 2. Extraction of Wolbachia genome in tissue: put 50 μL STE buffer containing mosquito tissue in a homogenizer, grind for about 45 seconds, add 2 μL protein kinase K (Proteinase K) (10 mg / mL ), place the reaction solution at 55°C for 30...

Embodiment 2

[0030] Embodiment 2: To the midgut, salivary gland, ovary, fat body and testis tissue of Anopheles stephensi (Anopheles stephensi) of artificial transfection B type Wolbachia (Wolbachia) Quantitative determination

[0031] 1. Acquisition of mosquito tissue samples containing Wolbachia: use CO 2 Anesthetize the mosquito to be tested (Anopheles stephenii artificially transfected with Wolbachia type B), and dissect and separate the mosquito tissue to be tested (midgut, salivary gland, ovary, fat body and testis) with a needle under a dissecting microscope ), the isolated tissue was rinsed three times with STE buffer and placed in 50 μL of STE buffer. The STE buffer solution belongs to the prior art buffer solution, and its formula is 100mM NaCl, 10mM Tris Cl, pH 8.0 and 1mM EDTA, pH 8.0.

[0032] 2. Extraction of Wolbachia genome in tissue: put 50 μL STE buffer containing mosquito tissue in a homogenizer, grind for about 45 seconds, add 2 μL protein kinase K (Proteinase K) (10 ...

Embodiment 3

[0038] Embodiment 3: Quantitative monitoring in different generation lines Wolbachia (Wolbachia) in the Anopheles stephensi (Anopheles stephensi) tissue distribution of artificial transfection B type Wolbachia (Wolbachia)

[0039] 1. Acquisition of mosquito tissue samples containing Wolbachia: use CO 2 Anesthetize the mosquitoes to be tested (Anopheles stepheni mosquitoes artificially transfected with type B Wolbachia (Wolbachia) of different generations), dissect and separate the mosquito tissues to be tested (midgut, salivary glands, ovaries, Fat body and testis), the isolated tissue was rinsed with STE buffer three times and placed in 50 μL of STE buffer. The STE buffer solution belongs to the prior art buffer solution, and its formula is 100mM NaCl, 10mM Tris Cl, pH 8.0 and 1mM EDTA, pH 8.0.

[0040] 2. Extraction of Wolbachia genome in tissue: put 50 μL of STE buffer containing mosquito tissue in a homogenizer, grind for about 45 seconds, add 2 μL of protein kinase K (Pr...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a primer for quickly and quantitatively detecting Wolbachia in tissues of a mosquito and a reagent box and a method thereof. According to the invention, different tissues containing Wolbachia are separated and obtained to extract Wolbachia genomic DNA; and then the Wolbachia DNA is subjected to Real-time PCR (Polymerase Chain Reaction) detection by using a specific PCR primer of Wolbachia. The realization of the invention is beneficial for quickly and quantitatively analyzing a large quantity of tissue samples containing Wolbachia, so that the distribution of Wolbachiain the tissues of the mosquito can be conveniently monitored in a large territorial scope on a large scale.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer for rapid quantitative detection of Wolbachia in mosquito body tissues, a kit and a method thereof. Background technique: [0002] Wolbachia is a symbiotic microorganism widely distributed in arthropods, and it may be the most abundant group of insect symbiotic microorganisms. It is distributed in Coleoptera, Diptera, Hemiptera, Homoptera, Hymenoptera, Lepidoptera and other insect species. It uses vertical transmission as its basic mode of transmission among host generations. It exists stably in the germ cells of the host, is transmitted to the offspring of the host through egg cells, and can regulate the reproductive activities of the host through various methods such as cytoplasmic incompatibility, feminization and andricide. These regulatory functions promote its widespread spread within the host population. [0003] Cytoplasmic incompatibility (CI) is the...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/01
Inventor 奚志勇朱俭
Owner GUANGZHOU WOLBAKI BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap