Application of lncRNAs as active tuberculosis specific markers

A tuberculosis and activity technology, applied in the field of molecular biology, can solve the problems of ineffective utilization of lncRNAs, complicated mechanism of action, and low prediction accuracy

Active Publication Date: 2018-02-02
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

LncRNAs promoters can also bind transcription factors, such as Oct3/4, Nanog, CREB, Sp1, c-myc, Sox2 and p53, and local chromatin histones also have characteristic modification methods and structural features, with low sequence conservation , only about 12% of lncRNAs can be found in other organisms other than humans, and its mechanism of action is very complex and has not yet been fully understood. The lncRNAs are not clear
In addition, some online websites such as the star

Method used

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  • Application of lncRNAs as active tuberculosis specific markers
  • Application of lncRNAs as active tuberculosis specific markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 The expressions of lnc-ABT1-3:5 and lnc-CCNT1-1:1 were significantly up-regulated in patients with active pulmonary tuberculosis

[0023] A1: Collect 20 patients with clinically confirmed active pulmonary tuberculosis and 20 healthy controls. Blood samples from all participants were drawn on an empty stomach in the morning, and 3.0 mL of peripheral blood was collected using disposable vacuum EDTA anticoagulant blood collection tubes, and mixed upside down; 3 times the volume of red blood cell lysate was added (300 μL of blood was added to 900 μL of red blood cell lysate), and mixed upside down. Mix well, put at room temperature for 5 minutes, invert several times during the period; centrifuge at 3000 rpm for 5 minutes, discard the supernatant. Suspend gently with 1 mL of erythrocyte lysate; centrifuge at 3000 rpm for 2 min, discard the supernatant; use 1 mL of TRIzol Reagent to suspend gently, and store in a -80°C refrigerator.

[0024] A2: Take out the TRIzo...

Embodiment 2

[0036] Example 2 Verification of the specificity of expression upregulation of lnc-ABT1-3:5 and lnc-CCNT1-1:1

[0037] A1: Collect 20 clinically confirmed pneumonia non-tuberculosis patients and 20 clinically confirmed lung cancer non-pulmonary tuberculosis patients. Blood samples from all participants were drawn on an empty stomach in the morning, and 3.0 mL of peripheral blood was collected using disposable vacuum EDTA anticoagulant blood collection tubes, mixed upside down; 3 times the volume of red blood cell lysate was added (300 μL of blood was added to 900 μL of red blood cell lysate), and inverted Mix well, place at room temperature for 5 minutes, invert several times during the period; centrifuge at 3000 rpm for 5 minutes, discard the supernatant. Suspend gently with 1 mL of erythrocyte lysate; centrifuge at 3000 rpm for 2 min, discard the supernatant; suspend gently with 1 mL of TRIzol Reagent, and store in a -80°C refrigerator.

[0038] A2: Take out the patient's T...

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Abstract

The invention discloses application of lncRNAs as active tuberculosis specific markers. Due to tissue specificity and spatial-temporal specificity of lncRNAs, different tissues are different in lncRNAs expression quantity, and the same tissue or organ varies in lncRNAs expression quantity in different growth stages. Two lncRNAs serving as the active tuberculosis specific markers are disclosed forthe first time, a whole blood sample which can be acquired easily, directly and clinically is combined with designed primer pairs to verify lncRNAs differential expression through RT-qPCR, and quickness, simplicity, time saving, high detection sensitivity and low sample consumption are realized. The method is quite suitable for active tuberculosis diagnosis by lncRNAs in differential expression, and important significances to researching of tuberculosis propagation control and reduction of the tuberculosis occurrence rate are achieved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for preparing lncRNAs specific to active pulmonary tuberculosis. Background technique [0002] Tuberculosis is a major global public health problem that seriously threatens human health. Pulmonary tuberculosis is the most common clinical type of tuberculosis. Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis infection. According to the World Health Organization (WHO), in 2015, there were about 10.4 million new cases of tuberculosis worldwide, 480,000 new cases of multidrug-resistant tuberculosis, and 1.4 million deaths from tuberculosis. The number of cases in China ranks second in the world. It is estimated that nearly 550 million people are infected with tuberculosis, and 1 million new cases of tuberculosis occur every year. The mortality rate was 2.32 / 100,000. Since tuberculosis is mainly airborne and has high morbidity a...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 张星朱敏杨荣胡小龙赵卫峰甘建和
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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