Reagent for mRNA detection and application
A technology of reagents and uses, applied in the field of mRNA detection reagents, can solve the problems of lack of early screening and rapid diagnosis, time-consuming, and inability to effectively distinguish active tuberculosis patients, and achieve the effect of accurate, fast and easy-to-operate tuberculosis diagnosis
Inactive Publication Date: 2017-09-22
SHENZHEN YOUSHENGKANG MEDICAL LAB CO LTD
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Problems solved by technology
However, there is currently a lack of effective early screening and rapid diagnosis methods
Sputum Mycobacterium tuberculosis microbiological examination has high specificity and is currently the gold standard for diagnosing active pulmonary tuberculosis. The sensitivity of genetic testing is quite different; the cellular immunological methods tuberculin skin test (TST) and tuberculosis interferon release assay (IGRA) are not effective in distinguishing active tuberculosis patients from those with latent tuberculosis infection
[0004] In view of the serious shortage of current detection technology for diagnosing active tuberculosis, which cannot meet the requirements of clinical and tuberculosis prevention and control, WHO put forward new technical recommendations for tuberculosis diagnostic products in December 2010: 1) In order to improve the detection accuracy, it is recommended to use non-sputum Fluid samples (such as blood) for testing
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example 1
[0064] In this embodiment, the crowd is divided into three groups: tuberculosis patients, latently infected people, and healthy people (20 cases each). By detecting GBP1 gene, DUSP3 gene, KLF2 gene, ID3 gene, P2RY14 gene, IFITM3 gene, GBP5 gene and other In tuberculosis patients, there was an obvious trend of up-regulation expression. The expression of KLRB1 gene was significantly down-regulated in tuberculosis patients.
[0065] In this example, quantitative RT-PCR method was used to detect the expression changes of GBP1 gene, DUSP3 gene, KLF2 gene, ID3 gene, P2RY14 gene, IFITM3 gene, GBP5 gene or KLRB1 gene. Specific steps are as follows
[0066] Step 1: Extraction of whole blood RNA;
[0067] RNA extraction was performed on blood samples stored in RNA stabilizer (product number: DP440) using Qiagene (product number: DP433). The specific operation is: take the blood sample stored in the RNA stabilizer and place it at room temperature, then centrifuge at 6600 rpm for 10 mi...
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The invention provides a reagent for mRNA detection and application. The reagent for mRNA detection comprises a gene primer and an internal reference primer. The application provided by the invention is characterized by preparing a product for distinguishing latent infection and active tuberculosis. The reagent has the beneficial technical effects that the product provided by the invention can be used for distinguishing active tuberculosis infection and latent infection, healthy people and other nontuberculous infectors, and has the detection sensitivity and the specificity being larger than 85 percent; has the detection sensitivity on patients with human immunodeficiency virus combined infection being larger than 80 percent; has the detection sensitivity on child patients with tubercle bacillus culture positive being larger than 70 percent. The operation can be carried out simply and conveniently, and the detection time is less than 8 hours.
Description
technical field [0001] The invention belongs to the technical field of detection, in particular to reagents and applications for mRNA detection. Background technique [0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis infection. According to the World Health Organization (WHO) estimates, in 2014, there were 9.6 million new cases of tuberculosis in the world, and about 1.5 million patients died of tuberculosis every year. At present, my country is the second largest country with a high burden of tuberculosis after India in the world. There are more than 1.2 million new active tuberculosis patients and 130,000 deaths in the country every year. Despite the availability of effective anti-TB drugs, tuberculosis is still the number one killer of infectious diseases. [0003] Due to the atypical symptoms of patients with combined diseases, the prevention and control of tuberculosis mainly depends on early detection and treatment. However...
Claims
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Patent Timeline
Login to View More IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6883C12Q2600/158C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 喻德华毛玉
Owner SHENZHEN YOUSHENGKANG MEDICAL LAB CO LTD