An oligonucleotide gene chip and its application in the detection of various germs

Abstract

The invention discloses an oligonucleotide gene chip and application of the oligonucleotide gene chip to the detection of various bacteria. Probes of the oligonucleotide gene chip can be hybridized with polymerase chain reaction (PCR) amplification products of M. tuberculosis, M. bovis, M. avium, M. paratuberculosis and Brucella respectively; and the probes have nucleotide sequences shown as SEQ ID No:25, SEQ ID No:31, SEQ ID No:35, SEQ ID No:37 and SEQ ID No:39 respectively. The gene chip is applied to the detection of the bacteria and has high repeatability, and interassay and intraassay coefficients of variation (CV) are less than 15 percent. The gene chip shows high accuracy on the detection of a pathogenic bacterium sample to be detected, which indicates that the method has high specificity and sensitivity; the high-throughput and parallel detection is realized; detection results are quick and accurate; the whole process from the preparation of nucleic acid to the finishing of the detection only needs 6 to 8h; and the gene chip has broad prospect in preparing 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 bacteria such as Mycobacterium bovis and Mycobacterium tuberculosis. health and safety. [0003] Bovine brucellosis (Brucellosis) is an infectious disease caused by bacteria such as Brucella abortus, which can lead to abortion of cows. At the same time, this 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 p...

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

[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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