Oligonucleotide gene chip and application of oligonucleotide gene chip to detection of various bacteria

Abstract

The invention relates to an oligonucleotide gene chip and its application in detecting various germs. The probes of the oligonucleotide gene chip can be combined with Mycobacterium tuberculosis (M.tuberculosis), Mycobacterium bovis (M.bovis), Mycobacterium avium (M.avium), Mycobacterium paratuberculosis ( M.paratuberculosis) and Brucella (Brucella) PCR amplification products were subjected to hybridization reaction; the probes had the nucleotides shown in SEQ ID NO: 25, SEQ ID NO: 31, SEQ ID NO: 35, SEQ ID NO: 37 and SEQ ID NO: 39 respectively sequence. The application of the gene chip in the invention to the detection of the pathogen has good repeatability, and the CV values ​​of the inter-batch and intra-batch variation coefficients are all less than 15%. The detection of pathogenic bacteria samples showed high accuracy, indicating that the method has good specificity and high sensitivity, and realizes high-throughput and parallel detection. The detection results are fast and accurate. From nucleic acid preparation to completion of detection, The whole process only takes 6-8 hours, and has broad prospects in the preparation of reagents and products for pathogen detection, import and export quarantine, and epidemiological analysis.

Description

technical field [0001] The invention belongs to the technical field of gene chips, and in particular relates to an oligonucleotide gene chip and its application in detecting various germs. Background technique [0002] Bovine tuberculosis (Bovis Tuberculosis) is an infectious disease caused by Mycobacterium bovis and Mycobacterium tuberculosis. Safety. [0003] Brucellosis is an infectious disease caused by bacteria such as Brucellosis abortus, which can lead to abortion of cows. At the same time, the bacteria can also infect humans and is difficult to cure. The number of cases of human infection with Brucella has been on the rise this year. , so the detection of this pathogen also has important public health significance. [0004] At present, the detection of tuberculosis pathogenic bacteria is mainly through the tuberculin test, but due to the influence of physical and biological factors, the detection rate of this type of pathogenic bacteria is not high; bacterial isola...

AI Technical Summary

Problems solved by technology

[0004] At present, the detection of tuberculosis pathogenic bacteria is mainly through the tuberculin test, but due to the influence of physical and biological factors, the detection rate of this type of pathogenic bacteria is not high; bacterial isolation and culture is a classic means of identification, but the growth cycle of mycobacteria is long , generally need to be cultured for 4 to 8 weeks, which is not conducive to rapid diagnosis; the establishment of PCR method can achieve rapid and sensitive detection, but it is easy to be contaminated and lead to false positive results

Mycobacterial infection can also be detected by detecting the level of IFN-γ released by peripheral blood lymphocytes, which has high sensitivity and specificity, but the disadvantage is that the cost is too high, and the detection must be completed within 8 hours after blood collection

For the detection of Brucellosis, test tube agglutination test and tiger red plate agglutination test are mainly used. This method will be affected by physical or biological factors, resulting in false positive or false negative results.

[0005] Therefore, it is urgent to develop a new, fast, accurate, and high-throughput detection technology for the above several pathogenic bacteria. The gene chip method has certain advantages due to its good specificity and high detection throughput. The related technology of pathogenic bacteria needs to be further studied

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