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High-flux loop-mediated isothermal amplification foodborne pathogen identification apparatus and detection method thereof

A ring-mediated isothermal and high-throughput technology, applied in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problems of increasing costs and increasing POC equipment, and achieve excellent performance, The effect of ingenious device design and small volume

Active Publication Date: 2016-10-12
ZHUHAI BAIRUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires complex and expensive equipment (optical fibers, LED sensors, etc.), and the cheapest real-time PCR analyzer also costs tens of thousands of yuan, which will increase the cost and increase the number of components of the POC equipment

Method used

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  • High-flux loop-mediated isothermal amplification foodborne pathogen identification apparatus and detection method thereof
  • High-flux loop-mediated isothermal amplification foodborne pathogen identification apparatus and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The principle diagram of the present invention is as figure 1 As shown, the high-throughput loop-mediated isothermal amplification device of the present invention for identifying foodborne pathogens includes a constant temperature mechanism 1, a reaction plate 2, a fluorescence excitation device 3, an image capture mechanism 4, and a portable computer 5, wherein the reaction plate 2 It is arranged on the top of the constant temperature mechanism 1 , the fluorescence excitation device 3 is arranged above the reaction plate 2 , the image pickup mechanism 4 is arranged above the fluorescence excitation device 3 , and the portable computer 5 and the image pickup mechanism 4 are connected through a data line 6 .

[0021] In this embodiment, the constant temperature mechanism includes a heat preservation chamber, a power supply, a heating element, a constant temperature controller, and a temperature sensor. The heating element and the temperature sensor are arranged in the hea...

Embodiment 2

[0048] The difference between this embodiment and embodiment 1 is that each stage of detection operation is specifically as follows:

[0049] Step 1: Prepare LMA Reagents and Buffers:

[0050] For each sample, a total volume of 10 μL of preparative loop-mediated isothermal amplification reagents and buffer solution preparation components is required;

[0051] Prepare loop-mediated isothermal amplification reagents and buffer reagents include the following components, the following content is the final concentration: betaine 500mM, deoxyribonucleotide triphosphate (dNTPs) mixture 1.3mM each, 1 × isothermal buffer (three Hydroxymethylaminomethane Hydrochloride (Tris-HCl) 20mM, Ammonium Sulfate 10mM, Triton X-100 8mM, Potassium Chloride 20mM), Magnesium Sulfate 5mM, Primer Mix 0.5μM, Bacillus stearothermophilus (BST) Hot start polymerase 0.4 unit / μL, fluorescent dye 20 μM;

[0052] Step 2: Preparation of the above samples Take appropriate clinical samples of food, patient secre...

Embodiment 3

[0056] The difference between this embodiment and embodiment 1 is that each stage of detection operation is specifically as follows:

[0057] Step 1: Prepare LMA Reagents and Buffers:

[0058] For each sample, a total volume of 50 μL of preparative loop-mediated isothermal amplification reagents and buffer solution preparation components is required;

[0059] Prepare loop-mediated isothermal amplification reagents and buffer reagents include the following components, the following content is the final concentration: betaine 1500mM, deoxyribonucleotide triphosphate (dNTPs) mixture 1.5mM, 1 × isothermal buffer (trihydroxy Methylaminomethane hydrochloride (Tris-HCl) 20mM, ammonium sulfate 10mM, Triton (Triton) X-100 8mM, potassium chloride 20mM), magnesium sulfate 15mM, primer mix 3.5μM, Bacillus stearothermophilus ( BST) hot start polymerase 1 unit / μL, fluorescent dye 20 μM;

[0060] Step 2: Preparation of the above samples Take appropriate clinical samples of food, patient se...

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Abstract

The invention relates to a high-flux loop-mediated isothermal amplification foodborne pathogen identification apparatus and a detection method thereof. The apparatus comprises an isothermal structure (1), a reaction plate (2), a fluorescence excitation device (3), an image shooting mechanism (4) and a portable computer (5), wherein the reaction plate (2) is arranged at the top of the isothermal structure (1), the fluorescence excitation device (3) is arranged above the reaction plate (2), the image shooting mechanism (4) is arranged above the fluorescence excitation device (3), and the portable computer (5) is connected with the image shooting mechanism (4). The above cheap image acquiring device connected with the portable computer is used to shoot the fluorescence intensity in the loop-mediated isothermal nucleic acid amplification process, and analysis is carried out to detect the foodborne pathogenic bacteria. The high-flux loop-mediated isothermal amplification foodborne pathogen identification apparatus has the advantages of small size, realization of battery power supply, and completion of high-flux nucleic acid identification of a sample through field work under commercial power-free conditions. The detection method is simple to operate, and is convenient and practical.

Description

technical field [0001] The invention is a high-throughput loop-mediated isothermal amplification identification device and detection method for food-borne pathogens, in particular, a high-throughput loop-mediated isothermal method for rapid identification of foodborne pathogens under POC conditions. The amplification identification device and detection method of foodborne pathogens belong to the innovative technology of high-throughput loop-mediated isothermal amplification identification device and detection method of foodborne pathogens. Background technique [0002] Medical diagnosis or care at the patient's side, known as point of care (POC), is an active research area with significant progress. There have been many reports in recent years. POC testing methods are currently available in primary care and emergency rooms. Some of the most common examples are one-time pregnancy tests and continuous glucose monitoring systems (CGMS). However, there are many other newer ap...

Claims

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Application Information

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IPC IPC(8): C12M1/38C12M1/34C12Q1/68
CPCC12Q1/6844C12Q2531/119C12Q2563/107C12Q2545/113
Inventor 刘和录张军李军涛侯敏阮承迈王海吴扬
Owner ZHUHAI BAIRUI BIOTECH
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