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Human immunodeficiency virus type 1 (HIV-1) nucleic acid quantitative detection kit

A human immunodeficiency, detection kit technology, applied in the biological field, can solve the problems of antibody decline, immune cross-reaction cycle, low sensitivity, etc.

Active Publication Date: 2019-04-05
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Serological neutralization tests and virus isolation and culture have disadvantages such as high safety risks, low sensitivity, immune cross-reaction, and long cycle, and are not suitable for primary screening of populations that require rapid and high-throughput
The antigen-antibody detection method has good sensitivity and rapid response. The obvious disadvantage is that there will be a "window period" of 1-3 months in the early stage of AIDS infection. Negative or uncertain, it is not conducive to the early diagnosis of HIV infection, resulting in missed detection, which may lead to the risk of HIV transmission; the end-stage antibody of AIDS also decreased significantly, which is not conducive to detection and tracking
Ordinary PCR has low sensitivity, HIV variability is very high, there are many popular subtypes, and the detection effect is not ideal

Method used

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  • Human immunodeficiency virus type 1 (HIV-1) nucleic acid quantitative detection kit

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0061] On the basis of comparing the nucleic acid sequences of HIV-1 known on GENBANK and the international HIV database, the specific conserved regions of the nucleic acid sequences of various subtypes were searched for, and the LTR regions and the distant LTR regions were selected. Two target nucleotide probes and corresponding primers were designed on the two conserved regions of the gene in the GAG ​​region. These primer probes are used in RT-PCR reaction enzyme systems containing thermostable DNA polymerase, reverse transcriptase, high-quality deoxyribonucleoside triphosphates (dNTPs) and Mg 2+ In the RT-PCR reaction solution with equal components, the circular amplification of nucleic acid in vitro is realized by a fluorescent PCR instrument.

[0062] According to the embodiment of the present invention, the main components of the kit involved in the present invention include: HIV-1 reaction solution A, HIV-1 reaction solution B, HIV-1 quantitative reference substance, n...

Embodiment 1

[0084] Embodiment 1: human immunodeficiency virus type 1 (HIV-1) nucleic acid quantitative detection kit (PCR-fluorescent probe method)

[0085] This kit uses fluorescent quantitative PCR method to quantitatively detect human immunodeficiency virus type 1 nucleic acid (HIV-1) RNA in serum or plasma samples, and is suitable for auxiliary diagnosis of human immunodeficiency virus type 1 infection and human immunodeficiency virus Monitoring the efficacy of drug therapy in people with type 1 infection.

[0086] Reagents included in the kit:

[0087]

[0088] Specific primer probe sequences are as above.

[0089] Inspection program:

[0090] 1. Sample processing and nucleic acid extraction (sample processing area)

[0091] The negative quality control, positive quality control, and quantitative reference in this kit are all involved in the extraction, and are used for monitoring the environment and quality control of PCR detection reagents. The internal standard solution in ...

Embodiment 2

[0128] Embodiment 2: Sensitivity detection experiment

[0129] Sensitivity reference samples come from a nominal concentration of 5×10 5 The national reference product of IU / ml (China Food and Drug Control Institute), with HIV-1 negative plasma diluted national reference product to 20IU / ml as the sensitivity reference product, totally 20 parts. Nucleic acid extraction was performed simultaneously with 20 sensitivity reference products, negative quality control products, HIV-1 strong positive quality control products, HIV-1 critical positive quality control products and 4 positive quantitative reference products.

[0130] The experiment was repeated 3 times in parallel and all obtained the same detection results, showing that the primers of the present invention have good specificity, the system has good amplification efficiency, and the sensitivity detection is good. The sensitivity of the kit of the present invention is 20IU / ml and repeated 20 times. Test, 3 The positive det...

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Abstract

The invention provides a human immunodeficiency virus type 1 (HIV-1) nucleic acid quantitative detection kit. Specifically, according to the detection kit provided by the invention, HIV-1 RNA nucleicacid is identified and quantified through a real-time fluorescent polymerase chain reaction technology. The detection kit can be widely applied in multiple fields of HIV-induced AIDS window detection,clinical diagnosis, scientific research, efficacy tracking, inspection and quarantine, plague prevention and control and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a quantitative detection kit for human immunodeficiency virus type 1 (HIV-1) nucleic acid. Background technique [0002] Human Immunodeficiency Virus (Human Immunodeficiency Virus; HIV), namely AIDS (AIDS, Acquired Immunodeficiency Syndrome) virus, is a lentivirus that infects cells of the human immune system and is a type of retrovirus. Human immunodeficiency virus was first identified in the United States in 1981. Because the HIV virus directly attacks the CD4-T lymphocyte system of the human body, the immune system is irreversibly damaged. Once the virus invades the cells of the body, it will integrate with the cells of the body and be difficult to eliminate for life. People with AIDS cannot be cured after being infected with various other diseases, and may develop malignant tumors, and the fatality rate is extremely high. On October 27, 2017, the list of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/6851C12Q1/703C12Q2600/16C12Q2600/166C12Q2531/113C12Q2521/107C12Q2537/143
Inventor 蒋析文夏乔李汉荣杨鸿辉廖丽丽王晶晶唐康明
Owner DAAN GENE CO LTD
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