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Molecular markers for detecting ciprofloxacin-resistant gene of broiler salmonellas and application thereof

A Salmonella, molecular marker technology, applied in the field of molecular biology, can solve the problems of improper drug use pressure, drug resistance, etc., and achieve high stability

Inactive Publication Date: 2017-10-03
SUZHOU JONATHAN NEW MATERIALS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Antibiotics are widely used in animal husbandry as the first-choice drugs for the prevention and treatment of bacterial diseases, but due to improper use and drug pressure, drug resistance has emerged

Method used

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  • Molecular markers for detecting ciprofloxacin-resistant gene of broiler salmonellas and application thereof
  • Molecular markers for detecting ciprofloxacin-resistant gene of broiler salmonellas and application thereof
  • Molecular markers for detecting ciprofloxacin-resistant gene of broiler salmonellas and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A molecular marker for detecting the ciprofloxacin resistance gene of broiler Salmonella, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.3-4; ID NO.5~6.

[0026] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0027] The application of the molecular marker in the preparation of a kit for detecting broiler Salmonella ciprofloxacin resistance gene.

[0028] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0029] The steps of the kit detecting broiler Salmonella ciprofloxacin resistance gene include:

[0030] (1) Sampling: select the purified broiler Salmonella, inoculate the culture medium by streaking to form a single colony, add 10 μL of 60% glycerol to a sterile EP tube, pick a single colony with an inoculation needle and wash it several times in glycerin, - Store...

Embodiment 2

[0040] A molecular marker for detecting the ciprofloxacin resistance gene of broiler Salmonella, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.3-4; ID NO.5~6.

[0041] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0042] The application of the molecular marker in the preparation of a kit for detecting broiler Salmonella ciprofloxacin resistance gene.

[0043] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0044] The steps of the kit detecting broiler Salmonella ciprofloxacin resistance gene include:

[0045] (1) Sampling: select the purified broiler Salmonella, streak and inoculate the medium to form a single colony, add 40 μL of 60% glycerol to the sterile EP tube, pick a single colony with an inoculation needle and wash it several times in glycerin, - Store at 20°C...

Embodiment 3

[0055] A molecular marker for detecting the ciprofloxacin drug-resistant gene of broiler Salmonella, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.3-4; P3, SEQ ID NO. ID NO.5~6.

[0056] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0057] The application of the molecular marker in the preparation of a kit for detecting broiler Salmonella ciprofloxacin resistance gene.

[0058] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0059] The steps of the kit detecting broiler Salmonella ciprofloxacin resistance gene include:

[0060] (1) Sampling: select the purified broiler Salmonella, inoculate the culture medium by streaking to form a single colony, add 60 μL of 60% glycerol to the sterile EP tube, pick a single colony with an inoculation needle and wash it several times ...

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Abstract

The invention discloses molecular markers for detecting a ciprofloxacin-resistant gene of broiler salmonellas and application thereof. The molecular markers have sequences of P1, SEQ ID NO. 1 to 2, P2, SEQ ID NO. 3 to 4 and P3, SEQ ID NO. 5 to 6. The molecular markers can fast detect carrying cases of salmonellas, provide reference for the relationship of salmonella resistance and a drug-resistant gene in a broiler slaughtering production line, provide the data basis for control of drug resistance to salmonellas in broiler slaughtering production and produce detection results having high stability.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a method for detecting microbial drug-resistant genes by means of molecular biology, in particular to a molecular marker for detecting broiler Salmonella ciprofloxacin drug-resistant genes and an application thereof. Background technique [0002] Antibiotics are widely used in animal husbandry as the first-choice drugs for the prevention and treatment of bacterial diseases, but due to improper use and drug pressure, drug resistance has emerged. Salmonella is a common zoonotic pathogen, which is very harmful to animals and humans. Salmonella has been constantly changing in the long-term evolution process, and multi-drug resistance has also appeared. Poultry is one of the main hosts of Salmonella, and poultry of various ages can be infected. As an important food-borne pathogen, Salmonella can infect humans through various channels such as contaminated livestock and poultry meat pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12N15/11C12R1/42
CPCC12Q1/689C12Q2600/106
Inventor 吕少波李苏杨李卓才
Owner SUZHOU JONATHAN NEW MATERIALS TECH