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A detection kit for mycobacterium tuberculosis (tb) nucleic acid

A Mycobacterium tuberculosis detection kit technology, applied in the field of biomedical clinical diagnostic reagents, can solve the problems of high biological risk, poor accuracy, and low sensitivity, and achieve the effects of good repeatability, easy operation, and high sensitivity

Active Publication Date: 2017-01-04
东北制药集团辽宁生物医药有限公司
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Problems solved by technology

[0006] The purpose of the present invention is to provide a Mycobacterium tuberculosis (TB) nucleic acid detection kit, which fundamentally solves the problems of low sensitivity, poor accuracy and high biological risk in the existing Mycobacterium tuberculosis (TB) nucleic acid PCR technology , the TaqMan fluorescent probe designed by it has the advantages of high sensitivity, strong specificity and good stability. The nucleic acid DNA extraction method is simple, the nucleic acid purity is high, and the cost is low. It is especially suitable for the detection of Mycobacterium tuberculosis (TB) and is easy to automate. and high-throughput detection

Method used

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  • A detection kit for mycobacterium tuberculosis (tb) nucleic acid
  • A detection kit for mycobacterium tuberculosis (tb) nucleic acid
  • A detection kit for mycobacterium tuberculosis (tb) nucleic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0023] (1) Example of fluorescent quantitative PCR detection of TB DNA

[0024] 1) Reagent preparation

[0025] a. According to the number of samples to be tested, negative control, critical positive control, and positive control, take the corresponding amount of magnetic bead suspension and TB inner Standard, fully mixed into a magnetic bead suspension-Mix for later use;

[0026] b. Preparation of PCR reaction solution

[0027] The composition of PCR buffer is 30mmol / L Tris-HCl (PH8.3), 10mmol / L(NH4)2SO4, 30mmol / LKCl, 4.5mmol / LMgCl2, 0.15mmol / L dNTPs. The concentration of the upstream primer and downstream primer for amplifying the target gene is 20 pmol, the concentration of the probe for target gene detection is 8 pmol, the concentration of the upstream primer and downstream primer of the internal standard is 8 pmol, and the concentration of the internal standard probe is 8 pmol.

[0028] c. According to the number of samples to be tested, negative control, critical posi...

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Abstract

A Mycobacterium tuberculosis (TB) nucleic acid detection kit used in the field of biomedical clinical diagnosis, the kit includes a magnetic bead nucleic acid extraction kit and a TB nucleic acid amplification kit, wherein the magnetic bead nucleic acid extraction kit includes Lysis binding solution, washing solution, eluent, magnetic bead solution; TB nucleic acid amplification kit includes TB-PCR reaction solution, enzyme mixture, TB-internal standard, negative quality control, borderline positive quality control, strong positive Quality control product: The TB-PCR reaction solution in the TB nucleic acid amplification kit includes probes for detecting target genes and internal standards, upstream primers and downstream primers for amplifying target genes and internal standards, and PCR buffer. The invention does not require high-speed centrifugation and boiling, simple and fast operation, low cost, low biological risk, and is convenient for automation and high-throughput detection; high detection sensitivity, strong specificity, good repeatability, and avoids false negative and false positive results; the introduction of high-efficiency The internal standard system can efficiently monitor the entire detection process.

Description

technical field [0001] The invention relates to a Mycobacterium tuberculosis (TB) nucleic acid detection kit in the field of biomedical clinical diagnostic reagents. Background technique [0002] Tuberculosis has always been one of the most serious public health problems in the world. There are 20 million people suffering from tuberculosis worldwide, with 8-10 million new cases every year, and about 3 million people die of tuberculosis every year worldwide. Tuberculosis, AIDS, and malaria are listed as the three major infectious disease killers affecting global health. The number of tuberculosis patients in my country ranks second in the world, with an annual incidence of about 1.3 million, accounting for 14.3% of the global incidence. The number of deaths from tuberculosis each year greatly exceeds the sum of deaths from other infectious diseases. On March 21, 2011, the Ministry of Health released the "Fifth National Tuberculosis Epidemiological Field Investigation Result...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 李望丰肖樊赵世源关尔鑫王丹蔡一荣曲俊杰董博赵雪楠
Owner 东北制药集团辽宁生物医药有限公司
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