Salmonella detection kit based on fimY gene and detection method thereof

A detection kit and technology for Salmonella, applied in biochemical equipment and methods, measurement/inspection of microorganisms, resistance to vector-borne diseases, etc., can solve problems such as operating procedures and unsatisfactory detection time specificity detection, and achieve specificity Strong, highly sensitive effects

Inactive Publication Date: 2011-08-03
SICHUAN AGRI UNIV
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Problems solved by technology

At present, the rapid detection methods of Salmonella mainly include immunoenzyme test, immunodiffusion method, latex agglutination test, po

Method used

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Embodiment Construction

[0022] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention are described in detail below.

[0023] Example Salmonella detection kit based on fimY gene and preparation method thereof

[0024] 1. Make the loop-mediated isothermal amplification kit for detecting Salmonella according to the following formula:

[0025] (1) Prepare LAMP reaction solution:

[0026] The 24 μl reaction system includes: the upstream outer primer F3 and the downstream outer primer B3 at a concentration of 0.2 μmol / L each, the upstream inner primer FIP and the downstream inner primer BIP at a concentration of 0.8 μmol / L each, and the upstream loop primer at a concentration of 0.4 μmol / L each. Primer LF and downstream loop primer LB, the concentration is 1.0 mmol / L deoxyribonucleotide triphosphate mixture, the concentration is 1mol / L betaine, the pH value is 8.8, the concentration is 25mmol / L tris hydroxymethyl ami...

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Abstract

The invention relates to the biological detection kit and method, in particular to a salmonella detection kit based on the fimY gene. The detection kit contains the following specific primers: a forward outer primer F3 shown in the SEQ ID NO.1, a reverse outer primer B3 shown in the SEQ ID NO.2, a forward inner primer FIP shown in the SEQ ID NO.3, a reverse inner primer BIP shown in the SEQ ID NO.4, a forward ring primer LF shown in the SEQ ID NO.5 and a reverse ring primer LB shown in the SEQ ID NO.6. The kit is prepared on the basis of the loop-mediated isothermal amplification technology, has high specificity and sensitivity and can detect salmonella easily and fast without using any expensive instrument, thus the kit is particularly suitable for the basic-level medical unit.

Description

technical field [0001] The invention relates to a biological detection kit and a method, in particular to a salmonella detection kit and a detection method based on the fimY gene. Background technique [0002] Salmonella (Salmonella) is a common zoonotic pathogen in Enterobacteriaceae. It can not only cause pullorum, piglet paratyphoid, abortion and other animal diseases, but also cause typhoid, paratyphoid, sepsis, gastroenteritis and food poisoning in humans. , in the food poisoning around the world, the poisoning cases caused by Salmonella rank the top. At present, the rapid detection methods of Salmonella mainly include immunoenzyme test, immunodiffusion method, latex agglutination test, polymerase chain reaction, immunofluorescence method and enzyme-linked immunosorbent assay (ELISA). Detection and other aspects are not ideal. [0003] Loop-mediated isothermal amplification (loop mediated isothermal amplification, LAMP) is a new nucleic acid detection method, which ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCY02A50/30
Inventor 程安春唐田汪铭书陈孝跃
Owner SICHUAN AGRI UNIV
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