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Brucella rapid detection kit based on mass-spectrometric technique

A Brucella and kit technology, applied in the field of protein fingerprint detection, can solve problems such as the complex composition of bacterial blood samples, achieve the effect of saving detection time and cost, and eliminating hazards to personnel and the environment

Inactive Publication Date: 2013-09-18
ICDC CHINA CDC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complex composition of bacterial blood samples, it cannot be directly detected by MLADI-TOF MS
At present, the identification of bacterial blood by MALDI-TOF MS relies on the front-end kit (MALDI Sepltyper kit, Bruker Company) to enrich pathogens and the back-end pathogen database to determine the mode. So far, there is no MALDI-TOF MS technology at home and abroad. There are technical reports on the direct detection of pathogens in body fluids, and there is no report on the direct detection of Brucella in body fluids

Method used

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  • Brucella rapid detection kit based on mass-spectrometric technique
  • Brucella rapid detection kit based on mass-spectrometric technique
  • Brucella rapid detection kit based on mass-spectrometric technique

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Effect test

Embodiment 1

[0034] The construction of embodiment 1 brucella mass spectrometry model

[0035] 1. Samples and instruments

[0036] Brucella International Standard Strain dog breed RM6-66, sheep breed 16M, cattle breed 544A and 32 cases of clinical blood samples. Among them, 3 international standard strains were used to construct the characteristic protein spectrum model of Brucella, and 32 cases of clinical blood were used as test samples. All strains have been confirmed using conventional Tb phage-specific lysis assays.

[0037] Matrix-assisted laser desorption time-of-flight mass spectrometry was Microflex LT from Bruker, USA. Data collection adopts Bruker’s FlexControlTM (version3.0) software to collect data, and Biotyper software performs spectrum optimization, reference spectrum construction and verification data search. The search parameters are: spectrum correction frequency critical value: 50; score calculation frequency critical value : 5; Maximum mass deviation of original spe...

Embodiment 2

[0049] The detection of embodiment 2 clinical actual samples

[0050] Sensitivity (sensitivity), also known as true positive rate (true positive rate), is actually the pathogen and is correctly judged as the percentage of the pathogen according to the standard of the detection method. Specificity, also known as true negative rate, is the percentage that is actually not the pathogen but is correctly judged as the pathogen according to the standard of the detection method.

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Abstract

The invention provides a kit for rapidly detecting brucella in a blood sample, relating to the technical field of fingerprint spectrum detection of proteins. According to the kit, based on the mass-spectrometric technique, a specific polypeptide mass-spectrometric model aiming at brucella detection in the blood is constructed, and a kit for carrying out the brucella detection by utilizing the mass-spectrometric technique is constructed. According to the kit provided by the invention, the brucella in the blood sample can be authenticated by utilizing a specific fingerprint spectrum, the accuracy is high, and the repeatability is good; the detection on 96 samples can be finished within 2 hours and can be finished at least 5 days in advance compared with the detection time in the conventional detection method; the detection cost is low, and the result is reliable, so that the kit has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of protein fingerprint detection, in particular to a mass spectrometry model for detecting Brucella characteristic proteins and a kit for detecting Brucella based on the mass spectrometry model. Background technique [0002] Brucella is a gram-negative intracellular parasite with no spores and no flagella and is pathogenic to humans and animals. It is the pathogen of brucellosis (brucellosis). As a serious zoonotic infectious disease, brucellosis has become one of the important public health problems in our country. The epidemic situation among humans and animals has risen sharply in the past 10 years, and the epidemic situation has shown a trend of spreading from pastoral and semi-pastoral areas to agricultural areas and even cities. The number of new cases is about 40,000 every year, and the prevention and control situation is very severe. The types of Brucella include sheep (3 types), cattle (8 types), p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 肖迪张建中姜海崔步云张慧芳
Owner ICDC CHINA CDC
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