Method and kit for quickly detecting interior label of salmonella by adopting reverse transcription fluorescent quantitation

A detection kit and Salmonella technology, applied in the field of food safety, can solve problems such as long time consumption, and achieve the effects of rapid detection method, high sensitivity and wide detection adaptability

Active Publication Date: 2012-09-19
SHANGHAI JIAO TONG UNIV +1
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional biochemical detection methods can distinguish dead bacteria from live bacteria, but it takes a long time; and using real-time fluorescent quantitative PCR (real-time PCR, referred to as RT-PCR) method, there is no effective method to eliminate false positives caused by inhibitors. Negative problems, so it is necessary to develop products that effectively monitor false negatives to avoid missed detections

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
  • Method and kit for quickly detecting interior label of salmonella by adopting reverse transcription fluorescent quantitation
  • Method and kit for quickly detecting interior label of salmonella by adopting reverse transcription fluorescent quantitation
  • Method and kit for quickly detecting interior label of salmonella by adopting reverse transcription fluorescent quantitation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0033] Hereinafter, the invention will be described through specific examples. If it is not specified in particular, it can be obtained according to "Molecular Colon Experiment Guide" (Third Edition) (Cold Spring Harbor Laboratory Press), "Cell Experiment Guide" (Science Press, Beijing, China, 2001), "RNA Experimental Technical Manual" (Science Press, Beijing, China, 2004), "Immunoassay Technology" (Science Press, Beijing, China, 1991) and other experimental manuals and references cited in this paper The methods listed are implemented.

[0034] 1.1 Main reagents and instruments

[0035] β-mercaptoethanol, 2.46mol / l, guanidine isothiocyanate extraction buffer, purchased from Shanghai Baihuishen Biotechnology Co., Ltd.; proteinase K, 20mg / ml, Tiangen Biotechnology (Beijing) Co., Ltd.; isopropyl Alcohol (Cat.No.80109218), ethanol (Cat.No.10009218), chloroform (Cat.No.10006818) were purchased from Shanghai Sinopharm Group; RNAprep pure cultured cells / bacterial total RNA extracti...

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 quick detection method by using an interior label and taking a sigD/sigE gene of salmonella as a target through reverse transcription fluorescent quantitation PCR (Polymerase Chain Reaction). In the quick detection method, an RNA (Ribonucleic Acid) interior label obtained by PCR amplification for three times is composed. In addition, the invention further provides a corresponding kit and application. The method disclosed by the invention can be widely applied to the fields such as research of laboratories, food safety and health and hygiene. The phenomenon of false negative missed detection can be avoided, so that the PCR amplification is effectively monitored; in addition, the detection specificity of salmonella is strong, so that dead bacteria and live bacteriacan be effectively distinguished; the quickness and high detection sensitivity are obtained; and the method can be applied to high-throughput screening of the salmonella in the food.

Description

technical field [0001] The invention belongs to a nucleic acid detection method in the field of food safety. Specifically, the present invention relates to a method, a kit, and an application for rapid detection of internal standard reverse transcription fluorescence quantitative detection of Salmonella sigD / sigE gene band. Background technique [0002] Salmonella (Salmonella spp.) is one of the important food-borne pathogens, which is mainly transmitted and infected through meat, milk, eggs and other foods, causing typhoid fever, sepsis, acute gastroenteritis and other diseases. Among the cases of bacterial food poisoning in various countries in the world, food poisoning cases caused by Salmonella often rank first or second. Establish an effective quantitative detection method for food-borne Salmonella, in order to improve the detection speed and accuracy of the bacteria, so that the contaminated food can be processed in time, and it is widely used in public health, food h...

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): C12Q1/68C12R1/42C12R1/89
Inventor 史贤明杨捷琳周敏潘良文杨柳
Owner SHANGHAI JIAO TONG UNIV
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