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Gene chip detection method for detecting tubercle bacillus infection from clinical specimen

A gene chip and detection method technology, applied in the field of gene chip detection for tuberculosis infection, can solve the problems of long culture time, low positive rate, poor specificity, etc., and achieve the effect of short time-consuming, high positive rate, and easy acquisition

Inactive Publication Date: 2014-02-12
HARBIN MEDICAL UNIVERSITY
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Problems solved by technology

The disadvantage of the CSF acid-fast bacilli smear method is that the positive rate is low, about 10%, and the bacteria in the sample must be ≥ 10%. 4 / ml can only be found, poor specificity
The shortcomings of the Mycobacterium tuberculosis culture detection method are that the operation is complicated, the culture time is long, it takes 4 to 8 weeks, and the positive rate is low, mostly in the range of 20 to 30%.

Method used

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  • Gene chip detection method for detecting tubercle bacillus infection from clinical specimen

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Embodiment Construction

[0015] Gene chip design and preparation:

[0016] 1. Gene chip design. Mycobacterium tuberculosis small RNA genes (target genes) include As1726, As1890, Aspks, F6, Asdes, G2, C8, B11 and B55 genes, the aforementioned genes are prior art. The gene chip includes 9 sets of probe sequences related to genes, and each set of probe sequences is complementary to 1 gene sequence of Mycobacterium tuberculosis small RNA. The gene chip also includes 1 positive control gene U6 for quantification and 2 irrelevant sequences as negative control.

[0017] 2. Gene chip preparation. Probes are designed according to the sequence of the target gene. Each group of probe sequences has 4-8 probes with a length of 14-50bp, which are sent to the reagent company for synthesis. Each probe is 20D.

[0018] Probes were dissolved in Tiris-EDTA buffer to a final concentration of 100 μM.

[0019] Mix 100uL probe with 300uL 0.2M PBS buffer, shake and mix at room temperature, ready for sample application. ...

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Abstract

The invention relates to a gene chip detection method for detecting tubercle bacillus infection from a clinical specimen. The gene chip detection method is used for detecting small RNA (Ribonucleic Acid) of tubercle bacillus in the clinical specimen. The method comprises the following steps: 1, extracting total RNA in the clinical specimen; 2, performing reverse transcription by taking the total RNA extracted in the step 1 to synthesize cDNA (complementary Deoxyribonucleic Acid) and marking; 3, detecting the marked cDNA synthesized in the step 2 by using a gene chip.

Description

technical field [0001] The invention relates to a gene chip detection method for detecting Mycobacterium tuberculosis infection from clinical specimens. Background technique [0002] In the prior art, commonly used detection methods for tuberculosis infection include CSF acid-fast bacilli smear method and Mycobacterium tuberculosis culture detection method. The disadvantage of the CSF acid-fast bacilli smear method is that the positive rate is low, about 10%, and the bacteria in the sample must be ≥ 10%. 4 / ml can only be found, and the specificity is poor. The shortcomings of the Mycobacterium tuberculosis culture detection method are complicated operation, long culture time, 4-8 weeks, and low positive rate, mostly 20-30%. Contents of the invention [0003] The object of the present invention is to provide a gene chip detection method for detecting Mycobacterium tuberculosis infection from clinical specimens, which is simple in operation, short in time consumption and ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6837C12Q2565/501
Inventor 张凤民李玉军付英梅翟爱霞赵吉子李辉
Owner HARBIN MEDICAL UNIVERSITY
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