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CAMP detection primer group of bacillus cereus enterotoxin gene and kit

A technology for Bacillus cereus and detection primers, which is applied to the determination/testing of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems that the detection technology of Bacillus cereus enterotoxin gene has not been reported, and achieve Achieve fast and accurate detection, shorten the detection time, and simplify the operation

Active Publication Date: 2019-09-20
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the CAMP detection technology of Bacillus cereus enterotoxin genes entFM and bceT

Method used

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  • CAMP detection primer group of bacillus cereus enterotoxin gene and kit
  • CAMP detection primer group of bacillus cereus enterotoxin gene and kit
  • CAMP detection primer group of bacillus cereus enterotoxin gene and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0036] A kit for detecting Bacillus cereus, comprising a CAMP detection primer set of the Bacillus cereus enterotoxin gene, said primer set including two sets of primers, respectively for amplification of the entFM region of the Bacillus cereus enterotoxin gene Primers (entFM-NF; entFM-NR) and primers (bceT-NF; bceT-NR) for amplifying the bceT region of the Bacillus cereus enterotoxin gene;

[0037] The kit includes several CAMP reaction tubes containing the CAMP working solution, which is prepared by putting the above primer set into DEPC-treated water; the kit also has a CAMP reaction tube containing the entFM primer set and a CAMP reaction tube containing the bceT primer set. CAMP reaction tube. The CAMP reaction tube containing the entFM primer set, in which the concentrations of entFM-NF ​​and entFM-NR are both 1.6 μM, and the volume of the working solution is 1 μL; the CAMP reaction tube containing the bceT primer set, in which the concentrations of bceT-NF and bceT-NR a...

Embodiment 2

[0052] The following reactions were carried out using the kit in Example 1.

[0053] Use the Eva Green chromogenic solution in the kit to verify the amplification reaction to the Bacillus cereus enterotoxin gene bceT.

[0054] The reaction solution system is 25 μL in total, and the combination is as follows:

[0055] 20mM Tris-HCl (pH8.8); 10mM KCl; 10mM (NH 4 ) 2 SO 4 ; 6mM MgSO 4 ; 0.1% Triton X-100; 1M betaine; 1.4mM dNTP; 8U Bst DNA polymerase; 1μL Eva green; 1μL nucleic acid sample;

[0056] Primers: 1 μL of CAMP working solution containing bceT primer set;

[0057] Target: Bacillus cereus-bceT dsDNA / GenBank: D17312.1;

[0058] At the same time, no target control group was set up.

[0059] The reaction conditions are as follows:

[0060] 1) adding the nucleic acid sample, CAMP reaction solution, and DEPC-treated water into the CAMP reaction tube containing the bceT primer set to obtain the amplification reaction solution;

[0061] 2) Set the Applied Biosystems Qu...

Embodiment 3

[0065] A kit for detecting Bacillus cereus, comprising a CAMP detection primer set of the Bacillus cereus enterotoxin gene, said primer set including two sets of primers, respectively for amplification of the entFM region of the Bacillus cereus enterotoxin gene Primers (entFM-NF; entFM-NR) and primers (bceT-NF; bceT-NR) for amplifying the bceT region of the Bacillus cereus enterotoxin gene;

[0066] The kit includes several CAMP reaction tubes containing the CAMP working solution, which is prepared by putting the above primers into DEPC-treated water; the kit also has a CAMP reaction tube containing the entFM primer set and a CAMP reaction tube containing the bceT primer set reaction tube. CAMP reaction tube containing entFM primer set, in which the concentrations of entFM-NF ​​and entFM-NR are both 2.0 μM, working solution volume 2.0 μL; CAMP reaction tube containing bceT primer set, in which the concentrations of bceT-NF and bceT-NR Both were 2.0 μM, and the volume of the w...

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Abstract

The invention relates to a CAMP detection primer group of a bacillus cereus enterotoxin gene and a kit. The primer group comprises two primers which are respectively a primer for amplifying the entFM region fragment of the bacillus cereus enterotoxin gene and a primer for amplifying the bceT region fragment of the bacillus cereus enterotoxin gene. The primer group provided by the invention is high in sensitivity and high in specificity, and the kit prepared by the primer group can quickly and accurately detect whether the sample contains bacillus cereus or not, and simultaneously judge whether the sample contains toxigenic genes entFM and bceT. Moreover, because the primer group provided by the invention has extremely high specificity, the time required for CAMP amplification is short, the detection time is further shortened, and operation is simplified.

Description

technical field [0001] The invention belongs to the technical field of bacterial gene detection, and in particular relates to a CAMP detection primer set and a kit for Bacillus cereus enterotoxin gene. Background technique [0002] Bacillus cereus is a non-encapsulated, motile, spore-producing, facultative aerobic, food-borne opportunistic pathogen, widely distributed in air, soil, sewage and various raw and cooked In food and food raw materials. After eating food contaminated by Bacillus cereus, people usually have vomiting or diarrhea, or both symptoms of poisoning within 8 to 16 hours. Vomiting type poisoning is mainly caused by vomitoxin, while diarrhea type is caused by various enterotoxins. Bacillus cereus poses a threat to food safety and human health and safety. Therefore, it is necessary to understand the potential toxicity of Bacillus cereus in food and establish a rapid and accurate detection program to monitor it. [0003] At present, the detection method of B...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/6844C12Q2531/119C12Q2537/1376Y02A50/30
Inventor 陈旭包海燕李薇石波乔宇彭晴岳喜庆乌日娜武俊瑞
Owner SHENYANG AGRI UNIV
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