Kit for detecting Salmonella spp.

A Salmonella and kit technology, applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, material analysis by observing the impact on chemical indicators, etc., can solve problems such as the lack of simple detection methods for Salmonella

Inactive Publication Date: 2010-09-08
天津朝海科技有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

There is currently a lack of simple, rapid, easy-to-use and low-cost detection methods for Salmonella

Method used

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  • Kit for detecting Salmonella spp.
  • Kit for detecting Salmonella spp.

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Embodiment Construction

[0015] The kit of the present invention includes 2 tubes of solution, one tube is 50 μl of nano gold particle solution labeled with a molecular probe that can recognize the specific sequence of Salmonella, and the other tube is 250 μl that can amplify the Salmonella DNA in the sample to be detected Or a loop-mediated isothermal nucleic acid amplification system that amplifies the Salmonella RNA in the sample to be detected after a reverse transcription process.

[0016] The gold nanoparticle solution contains gold nanoparticles with a diameter of 20nm at a concentration of 1.2nM, and the surface of the gold nanoparticles is marked with a sulfhydryl-modified DNA probe.

[0017] The components of the loop-mediated isothermal nucleic acid amplification system are as follows: 0.2 μM F3 and B3 primers, 1.6 μM FIP and BIP primers, 0.4M betaine (Sigma-Aldrich, St.Louis, MO), 10 mM MgSO 4 , 1.4mM dNTPs, 1×ThermoPol reactionbuffer (New England Biolabs, Ipswich, MA), 80 U Bst DNA polyme...

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Abstract

The invention relates to a kit for detecting product DNA of Salmonella spp. DNA amplified through a loop-mediated isothermal nucleic acid amplification technology by using gold nanoparticles, belonging to the technical field of biological detection. In combination with the loop-mediated isothermal nucleic acid amplification technology and a biological nanotechnology, the invention has the advantages that the sensitivity of biological molecular detection is improved, the operation of the method is simple, the detection is rapid, the accuracy is high, special instruments and devices are not required, and the kit can be widely used for high-sensitivity Salmonella spp. detection in fields such as family diagnosis, clinical diagnosis, infectious disease control, environmental monitoring, inspection and quarantine, biotechnology and the like. The kit comprises: (1) gold nanoparticles labeled with molecular probes capable of identifying specific sequences of Salmonella spp.; and (2) a loop-mediated isothermal nucleic acid amplification system capable of amplifying Salmonella spp. DNA in a sample to be detected or capable of amplifying Salmonella spp. RNA in the sample to be detected after reverse transcription.

Description

1. Technical field [0001] The invention relates to a kit for detecting the DNA of Salmonella DNA or RNA amplified by a loop-mediated isothermal nucleic acid amplification technology by using nano-gold particles, and belongs to the technical field of biological detection. 2. Background technology [0002] The high sensitivity, high specificity, simple and rapid detection of genes (DNA) and their transcription products (RNA) is very useful for early diagnosis and treatment of diseases, as well as in the fields of health and epidemic prevention, environmental monitoring, inspection and quarantine, etc. Significance. In the past, more accurate and sensitive detection methods in this regard include fluorescent labeling, radioactive labeling and other methods. These methods require complex operations and special equipment, and their application range is very narrow. In 1993, Dr.Kary B.Mullis of PE Company invented the polymerase chain reaction (Polymerase Chain Reaction), referre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10G01N21/78
CPCY02A50/30
Inventor 孙国明
Owner 天津朝海科技有限公司
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