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Qualitative and quantitative detection method for bacterium AB5 enterotoxin proteins

A quantitative detection method and detection method technology, applied in the field of membrane electrospray ionization mass spectrometry, can solve problems such as complex processing methods, and achieve the effects of improving detection sensitivity, rapid detection, and saving desalination steps

Inactive Publication Date: 2017-06-13
ICDC CHINA CDC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the problem with the current method is that the sample pretreatment method is still relatively complicated, and it cannot meet the requirements of fast qualitative and quantitative detection of AB on the premise of ensuring high sensitivity. 5 Enterotoxin detection

Method used

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  • Qualitative and quantitative detection method for bacterium AB5 enterotoxin proteins
  • Qualitative and quantitative detection method for bacterium AB5 enterotoxin proteins
  • Qualitative and quantitative detection method for bacterium AB5 enterotoxin proteins

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Embodiment Construction

[0046] Embodiments of the present invention are described below with reference to the drawings. Elements and features described in one drawing or one embodiment of the present invention may be combined with elements and features shown in one or more other drawings or embodiments. It should be noted that representation and description of components or processes that are not relevant to the present invention and known to those of ordinary skill in the art are omitted from the drawings and descriptions for the purpose of clarity.

[0047] The present invention will be further described below in conjunction with the accompanying drawings.

[0048] 1 Bacterial strains and reagents used in the detection of the present invention

[0049] 1.1 Strains (Table 1)

[0050] Table 1 strain list

[0051]

[0052]

[0053] a ND:AB 5 Enterotoxin negative

[0054] 1.2 Reagents (Table 2)

[0055] Table 2 Reagent list

[0056]

[0057]

[0058] 2 Experimental methods

[0059]...

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Abstract

The invention relates to a qualitative and quantitative detection method for bacterium AB5 enterotoxin proteins. The method comprises the following steps: extracting a protein in a to-be-detected sample by adopting ultrafiltration so as to obtain a sample protein; performing enzyme digestion on the sample protein; detecting AB5 enterotoxin specific peptide fragments by adopting membrane electrospray ionization mass spectrometry, and qualitatively and quantitatively determining existence of AB5 enterotoxin coding genes on the peptide fragment level, wherein the AB5 enterotoxins respectively refer to Ctx, LT, Stx1 and Stx2 of which sequences are respectively shown as SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.5 and SEQ ID NO.7. With the adoption of the membrane electrospray ionization mass spectrometry, matrix effects caused by small molecules introduced by enzyme digestion are effectively removed, and a desalting step in the conventional mass spectrometry process is saved, so that the method disclosed by the invention can realize rapid detection at extremely high sensitivity.

Description

technical field [0001] The present invention relates to the application of membrane electrospray ionization mass spectrometry, specifically, relates to a kind of application membrane electrospray ionization mass spectrometry to bacterial AB 5 Qualitative and quantitative detection methods of enterotoxin protein. Background technique [0002] Worldwide, enteric infections continue to play an important role in contributing to the disease burden. The resulting reduction in productivity, decline in quality of life, and increase in medical expenses have significantly affected economic development. can generate AB 5 Bacterial pathogens of enterotoxins play an important role in intestinal infections. [1] AB 5 Enterotoxin gets its name because it contains two subunits, A and B, in which the catalytic A subunit can interfere with the necessary physiological functions of the host, while the B subunit is responsible for regulating the recognition of host receptors. [2] [0003] A...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N27/68
CPCG01N27/68G01N33/6848
Inventor 张玫阚飙徐建国濮吉滕中秋付秀萍
Owner ICDC CHINA CDC
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