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Enterobacter sakazakii detection reagent and preparation method thereof

A technology of Enterobacter sakazakii and detection reagents, which is applied in the field of bacterial detection reagents and its preparation, can solve the problems that colloidal gold test strips cannot be quantitatively detected, and achieve the effects of large detection linear range, fast detection, and convenient use

Inactive Publication Date: 2015-06-10
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention aims to solve the problem that the existing colloidal gold test strips for detecting Enterobacter sakazakii cannot be quantitatively detected, and provides a detection reagent for Enterobacter sakazakii and its preparation method

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  • Enterobacter sakazakii detection reagent and preparation method thereof
  • Enterobacter sakazakii detection reagent and preparation method thereof
  • Enterobacter sakazakii detection reagent and preparation method thereof

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specific Embodiment approach 1

[0019] Specific embodiment 1: In this embodiment, the detection reagent of Enterobacter sakazakii is composed of reagent A and reagent B, wherein the reagent A includes a buffer, a coagulant, BSA and a preservative; Colloidal gold particles, stabilizers, buffers and preservatives for Enterobacter polyclonal antibodies; the concentration of the buffer in reagent A is 20-50mmol / L, the concentration of the coagulant is 2% (w / v), and the concentration of BSA The concentration of the preservative is 0.1% to 0.5% (w / v); the concentration of the buffer in reagent B is 20 to 50mmol / L, and the polyclonal antibody labeled with anti-Enterobacter sakazakii The concentration of the colloidal gold particles is 0.08mg~0.8mg / mL, the concentration of the stabilizer is 2%~5% (w / v), the concentration of the preservative is 0.1%~0.5% (w / v), and the reagent A The volume ratio of reagent B and reagent B is 5:1.

specific Embodiment approach 2

[0020] Specific embodiment two: the difference between this embodiment and specific embodiment one is that the buffers in reagent A and reagent B are both Tris-HCl buffer and HEPES buffer containing 0.5% to 5.0% (w / v) BSA , borate buffer, glycine buffer, or a mixture of several in any ratio. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0021] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the coagulant in reagent A is PEG6000 or Brij-35. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses enterobacter sakazakii detection reagent and a preparation method thereof and relates to bacterial detection reagent and a preparation method thereof. The enterobacter sakazakii detection reagent and the preparation method thereof are aimed to solve the problem that an existing colloidal gold test strip for detecting the enterobacter sakazakii cannot perform quantitative detection. The detection reagent is composed of reagent A and reagent B, and the reagent A comprises buffer solution, coagulation accelerator, BSA and preservative; the reagent B comprises colloidal gold particles labeled with anti-enterobacter sakazakii polyclonal antibody, stabilizing agent, buffer solution and preservative. The preparation method includes that firstly, adding the coagulation accelerator, BSA and preservative to the buffer solution to obtain the reagent A; secondly, mixing the stabilizing agent, preservative and buffer solution to obtain the mixed solution, and mixing the mixed solution with the colloidal gold particles labeled with anti-enterobacter sakazakii polyclonal antibody to obtain the reagent B; thirdly, mixing the reagent A with the reagent B to obtain the enterobacter sakazakii detection reagent. The enterobacter sakazakii detection reagent is large in detection linearity range, capable of performing quantitative detection, high in analysis sensitivity and convenient to use. The enterobacter sakazakii detection reagent is used for detecting the enterobacter sakazakii in milk or milk powder.

Description

technical field [0001] The invention relates to a bacteria detection reagent and a preparation method thereof, in particular to a reagent for detecting Enterobacter sakazakii in milk powder or milk and a preparation method thereof. Background technique [0002] Cronobacter sakazakii is a Gram-negative, motile, non-spore-forming and facultatively anaerobic. Previously considered to be Enterobacter cloacae (Enterobacter cloacae) until 1980 was renamed "Enterobacter sakazakii". In 2008, Enterobacter sakazakii was defined as a new genus of Enterobacter—Cronobacter spp. [0003] C. sakazakii is a pathogenic bacterium that can parasitize in animals and humans. It can cause severe neonatal meningitis, necrotizing enterocolitis and bacteremia, and can cause neurological sequelae and even death. The mortality rate caused by this bacterial infection is as high as 50%. In recent years, studies on C. sakazakii in food, beverages, processing raw materials, and ecological environment h...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/56916G01N33/531G01N2333/265
Inventor 孟利刘峰李梦洋李冲伟任洪波金海涛
Owner HEILONGJIANG UNIV