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Method for rapidly detecting S.aures by combining bacteriophage lyase with BLI

A phage lyase and lyase technology are applied in the field of rapid detection of S. aureus by phage lyase combined with BLI, which can solve the problems of complicated operation, many steps, and long time, and achieve the effect of simple operation, short time, and elimination of potential interference.

Pending Publication Date: 2022-02-11
JILIN UNIV
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Problems solved by technology

[0005] In order to solve the above technical problems, the present invention provides a method for rapid detection of S.aureus by combining phage lysing enzyme LysGH15 with BLI, so as to solve the problems of many steps, cumbersome operation and long time-consuming in the traditional detection method of S.aureus

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  • Method for rapidly detecting S.aures by combining bacteriophage lyase with BLI
  • Method for rapidly detecting S.aures by combining bacteriophage lyase with BLI
  • Method for rapidly detecting S.aures by combining bacteriophage lyase with BLI

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Embodiment

[0047] as attached Figure 1-6 Shown:

[0048] The invention provides a method for rapidly detecting S.aureus by combining phage lyase LysGH15 with BLI, comprising the following steps:

[0049] 1) Sensor activation: After pre-wetting the end of the AR2G sensor in distilled water for at least ten minutes, immerse in 40 mM 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and 20 mM N -Activate in the mixed activation reagent of hydroxysuccinimide for 150-450 seconds. When the specific activation time is set to 200-300 seconds, the carboxyl group modified on the surface of the sensor is efficiently activated to ensure the effective immobilization of subsequent phage lyase;

[0050]2) Immobilization of lyase: immerse the activated AR2G sensor in 12.5-100 μg / mL mutated LysGH15 diluted with acetic acid-sodium acetate buffer solution at pH 4-6 for 600-900 seconds, specifically During operation, immerse the activated AR2G sensor in 50-100 μg / mL mutated LysGH15 diluted with ...

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Abstract

The invention provides a method for rapidly detecting S.aures by combining bacteriophage lyase with BLI, relates to the technical field of staphylococcus aureus detection, and solves the problems of multiple steps, complicated operation and long time consumption of a traditional S.aures detection method. The method comprises the following steps: (1) activating a sensor, (2) immobilizing lyase, (3) sealing the AR2G sensor, and (4) detecting a sample and judging a result. The method has the advantages of simple operation, real-time monitoring, no mark and easily available result; the sample is detected at any time, target bacteria in the sample can be recognized and detected in a short time, and the pure sample detection time can be controlled within tens of seconds to dozens of minutes according to different concentrations of staphylococcus aureus contained in the sample; the lyase can specifically recognize target bacteria, so that the specificity of the method is ensured, and potential interference of non-target bacteria is eliminated; dead and viable bacteria can be distinguished according to whether the signal curve has a peak or not; the lyase is easy to obtain and convenient to prepare.

Description

technical field [0001] The invention belongs to the technical field of staphylococcus aureus detection, and more specifically relates to a method for rapidly detecting S. aureus with phage lyase combined with BLI. Background technique [0002] Staphylococcus aureus (S.aureus) is a Gram-positive bacterium ubiquitous in nature. It is a common foodborne pathogen and has potential hazards to the food processing industry and human health. In addition, it has low nutritional requirements and is highly resistant to adverse environments that can cause infectious diseases in humans and animals. Therefore, using a rapid and accurate detection method to detect S.aureus can greatly reduce the occurrence of food poisoning incidents. [0003] The traditional S.aureus detection method has the disadvantages of long cycle and complicated operation, which is difficult to meet the actual needs. With the continuous development of molecular biology techniques, a variety of nucleic acid-based d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/31
CPCG01N33/56938G01N21/31G01N2333/31
Inventor 顾敬敏刘潇张晓光韩文瑜
Owner JILIN UNIV
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