A set of peripheral blood gene markers for noninvasive diagnosis of active pulmonary tuberculosis
A technology of active pulmonary tuberculosis and markers, applied in the direction of recombinant DNA technology, biochemical equipment and methods, and microbial measurement/testing, can solve the problem of difficult to distinguish lung infection diseases, low sensitivity of sputum tuberculosis, correct Diagnosis delays and other issues, to achieve the effect of high accuracy, good stability, and convenient detection
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Embodiment 1
[0036] Example 1: A group of screening of peripheral blood gene markers for tuberculosis diagnosis comprising 56 genes
[0037] In order to screen the diagnostic markers for pulmonary tuberculosis, the present invention firstly analyzes the whole gene expression data in the peripheral circulating blood of a population used to discover the diagnostic markers for pulmonary tuberculosis. Among them, the population used to discover gene markers included 38 normal people, 8 patients with pneumonia, 8 patients with lung cancer and 16 patients with tuberculosis.
[0038] First, the present invention compares the complete gene expression in the peripheral blood of tuberculosis patients and normal people, screens out the differentially expressed genes in the peripheral blood of tuberculosis patients and normal people, and uses the SAM algorithm to realize the difference analysis of gene expression in the two populations. And use the q value method to control the false positive rate (Fa...
Embodiment 2
[0049] Example 2: Validation of a set of 56-gene gene markers in peripheral blood for the diagnosis of pulmonary tuberculosis in an independent population
[0050] The present invention obtains the gene expression data of all 56 gene markers in the peripheral blood of individuals from other groups of people to verify that the peripheral blood gene markers comprising 56 genes can distinguish tuberculosis patients from non-tuberculosis individuals, active tuberculosis patients and latent tuberculosis patients. Effectiveness in tuberculosis-infected individuals, tuberculosis patients and pneumonia patients, and tuberculosis patients and lung cancer patients.
[0051] The BL population (GEO number: GSE42826), the CA population (GEO number: GSE54992), the BE population (GEO number: GSE19491) were downloaded from the public database GEO (http: / / www.ncbi.nih.nlm.gov / geo) The expression data of all genes in the peripheral blood of individuals related to the AN population (GEO number: ...
Embodiment 3
[0077] Embodiment 3: Tuberculosis diagnostic kit based on 56 gene markers
[0078] Based on the 56 gene markers identified and verified by the present invention for the diagnosis of peripheral blood tuberculosis, the forward and reverse primers for amplifying each gene are designed, as shown in the sequence table SEQ ID NO.1-SEQ ID NO.112 shown, where:
[0079] SEQ ID NO.1 and SEQ ID NO.2 are forward primer and reverse primer of ALDH1A1 respectively;
[0080] SEQ ID NO.3 and SEQ ID NO.4 are the forward primer and reverse primer of ANKRD22 respectively;
[0081] SEQ ID NO.5 and SEQ ID NO.6 are forward primer and reverse primer of APOL2 respectively;
[0082] SEQ ID NO.7 and SEQ ID NO.8 are forward primer and reverse primer of BTN3A1 respectively;
[0083] SEQ ID NO.9 and SEQ ID NO.10 are CD74 forward primer and reverse primer respectively;
[0084] SEQ ID NO.11 and SEQ ID NO.12 are forward primer and reverse primer of DDX60 respectively;
[0085] SEQ ID NO.13 and SEQ ID NO...
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