Application of novel mixed matrix in MALDI-MS (matrix-assisted laser desorption ionization-mass spectrometry) bacterial identification

A mixed matrix, bacterial identification technology, applied in the direction of resisting vector-borne diseases, instruments, measuring devices, etc., can solve the problems of inability to distinguish similar bacteria, incomplete fingerprints, etc., and achieve accurate and highly sensitive identification of clinical drug delivery. , the effect of expanding the number of mass spectral peaks

Active Publication Date: 2021-11-02
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the embodiments of the present invention is to provide the application of a new mixed matrix in the identification of bacteria by MALDI-MS, aiming to solve the problem that the fingerprint spectrum obtained when the traditional matrix detects bacteria is not comprehensive enough and cannot distinguish similar bacteria

Method used

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  • Application of novel mixed matrix in MALDI-MS (matrix-assisted laser desorption ionization-mass spectrometry) bacterial identification
  • Application of novel mixed matrix in MALDI-MS (matrix-assisted laser desorption ionization-mass spectrometry) bacterial identification
  • Application of novel mixed matrix in MALDI-MS (matrix-assisted laser desorption ionization-mass spectrometry) bacterial identification

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Experimental program
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Embodiment 1

[0077] The step of detecting bacterial mixed matrix in MALDI mass spectrometry as follows:

[0078] Step a: cultured on solid medium were Pseudomonas aeruginosa and Enterobacter 24h;

[0079] Step 2: CHCA and CHCA-C3 were formulated in 50% acetonitrile containing 2.5% trifluoroacetic acid solution in a concentration of 10-15mg / mlCHCA CHCA-C3 substrate solution and stored at 4 ℃ refrigerator to standby;

[0080] Step Three: CHCA matrix solution were taken and CHCA-C3 substrate solution at a volume ratio of 1: 2 were uniformly mixed;

[0081] Step Four: a culture from step Enterobacter and Pseudomonas aeruginosa were obtained uniformly coated on a supporting MALDI-MS target plate;

[0082] Step Five: Enterobacter and Pseudomonas aeruginosa on Pseudomonas Step Four target plates after air-drying, the mixed matrix solution obtained in step three points each were taken 0.5-1μL Pseudomonas aeruginosa and Enterobacter on the dry ;

[0083] Step Six: A solution of step 5 was added dropw...

Embodiment 2

[0087] Mixed matrix determination step Shigella and E. coli in MALDI mass spectrometry as follows:

[0088] Step a: On solid medium were cultivated Escherichia coli and Shigella 24h;

[0089] Step 2: CHCA and CHCA-C3 were formulated in 50% acetonitrile containing 2.5% trifluoroacetic acid solution in a concentration of 10-15mg / mlCHCA CHCA-C3 substrate solution and stored at 4 ℃ refrigerator to standby;

[0090] Step Three: CHCA matrix solution were taken and CHCA-C3 substrate solution at a volume ratio of 1: 2 were uniformly mixed;

[0091] Step Four: obtained from step a culture of Escherichia coli and Shigella are uniformly coated on a supporting MALDI-MS target plate;

[0092] Step Five: after Step Four target plates on Escherichia coli and Shigella air drying, the mixture obtained in step three matrix solution were taken on 0.5-1μL point after dry Escherichia coli and Shigella;

[0093] Step Six: Escherichia coli step 5 was added dropwise a mixed solution of the matrix and S...

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Abstract

The invention is applicable to the technical field of bacterium identification, and provides application of a novel mixed matrix in MALDI-MS (matrix-assisted laser desorption ionization-mass spectrometry) bacterium identification, which comprises the following steps: 1, culturing to-be-detected bacteria on a solid culture medium for 24 hours; 2, respectively dissolving CHCA and CHCA-C3 in a solution containing 50% acetonitrile and 2.5% trifluoroacetic acid to prepare CHCA and CHCA-C3 matrix solutions; 3, respectively taking CHCA matrix liquid and CHCA-C3 matrix liquid, and uniformly mixing the CHCA matrix liquid and the CHCA-C3 matrix liquid according to a volume ratio of 1: 2; 4, uniformly coating the bacteria cultured in the step (1) on an MALDI-MS target plate; 5, after the bacteria on the target plate in the step 4 are air-dried, the mixed matrix solution obtained in the step 3 is dripped on the dried bacteria; 6, after the solvent in the step 5 is volatilized and crystallized, directly carrying out MALDI analysis. The novel MALDI mixed matrix can detect hydrophilic protein and hydrophobic protein at the same time, so that the number of mass spectrum peaks of an analyte is increased, bacteria can be identified more accurately, rapid and high-sensitivity distinguishing and identification of the bacteria are realized, and the accuracy of distinguishing the bacteria by using the MALDI-MS is improved.

Description

Technical field [0001] The present invention belongs to the technical field of bacterial identification, in particular, the application relates to novel mixed matrix in MALDI-MS in bacterial identification. Background technique [0002] Matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) has been widely applied to proteomics, metabolomics in clinical and microbiological analysis. It is a fast, sensitive and high-throughput analytical methods. Its principle is that the laser energy absorbed by a special small molecule organic compound as a substrate, protonation transfer, thereby enabling the analyte ions, thereby to achieve detection purposes. [0003] Traditional bacterial detection matrix fingerprint spectrum obtained is mostly hydrophilic proteins, which results in a conventional MALDI detection, can not distinguish between similar bacteria. But although many similarities to bacteria similar to the early clinical manifestations of the disease but are quite...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64
CPCG01N27/64Y02A50/30
Inventor 国新华赵楠
Owner JILIN UNIV
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