HIV-1 genotype and drug resistant mutation site detection kit and application thereof

A drug-resistant mutation site, HIV-1 technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Expensive testing kits and other issues

Inactive Publication Date: 2015-09-30
CAPITALBIO CORP +2
View PDF1 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these two detection systems are mainly established for the subtype B virus strains prevalent in Europe and the United States. Antimicrobial resistance detection of serotypes and other popular recombinant virus strains is not ideal
In addition, these two commercial drug resistance detection kits are expensive, which is not conducive to large-scale clinical application in my count

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • HIV-1 genotype and drug resistant mutation site detection kit and application thereof
  • HIV-1 genotype and drug resistant mutation site detection kit and application thereof
  • HIV-1 genotype and drug resistant mutation site detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Amplification and sequencing primers of the Pol gene region of embodiment 1, HIV-1

[0089] The present invention downloads the full-length genome sequence of HIV-1 virus worldwide from the HIV database (http: / / www.hiv.lanl.gov / ), and utilizes appropriate evolutionary analysis software to analyze the global, Asian and Chinese The evolutionary analysis of the full-length genome sequence of the HIV-1 virus in China was carried out, and the evolutionary relationship and trend of different subtypes of HIV-1 viruses were obtained. According to the evolutionary relationship, the full-length genome sequences of 13 subtypes, including B, B' (Thailand B), CRF-01AE, CRF-07BC, and CRF-08BC, with high prevalence and wide geographical coverage were selected, and the appropriate nucleic acid homology analysis was used The software carries out conservation analysis to the Pol gene region of HIV-1, finds out the highly conserved section of the candidate (conservation is greater than 98...

Embodiment 2

[0102] Embodiment 2, HIV-1 genotype drug resistance detection kit and detection method thereof

[0103] The kit of the present invention comprises RT-PCR amplification reagent A, RT-PCR amplification reagent B containing primer 1 and primer 2 in Example 1, RT-PCR amplification reagent C, and primer 3 in Example 1 PCR amplification reagent D with primer 8, sequencing amplification reagent 1-6 containing primer 3-primer 8 in Example 1. The detection method of the kit is as follows:

[0104] 1. Extraction of RNA

[0105] Extract the RNA in HIV-1 from the serum or plasma of the HIV-1 patient to be tested (from the AIDS Research Institute of the First Affiliated Hospital of China Medical University, informed by the patient, clinically diagnosed, containing drug resistance sites I84V, A71V and K101E) ,Specific steps are as follows:

[0106] 1. Add 20 μL proteinase K to a clean 1.5mL centrifuge tube;

[0107] 2. Add 200 μL of patient plasma sample to the centrifuge tube;

[0108...

Embodiment 3

[0186] Embodiment 3, the application of the detection kit of HIV-1 genotype drug resistance

[0187] Plasma samples of 50 HIV-1 infected persons with known genotypes and drug-resistant loci were collected (the First Affiliated Hospital of China Medical University, patients were informed and clinically diagnosed, and the genotypes and drug-resistant loci of 50 clinical samples were collected. Detecting result is shown in table 9), with the detection kit of HIV-1 genotype drug resistance and detection method thereof among the embodiment 2 to the genotype of 50 routine HIV-1 infected persons plasma samples and drug-resistant locus detection.

[0188] The genotype drug resistance detection results of 50 cases of clinical samples are as shown in Table 9: in 50 cases of clinical samples, the proportion of samples with drug resistance sites in the protease gene region was 22% (11 / 50); there was drug resistance in the reverse transcriptase gene region The sample ratio of the site is ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an HIV-1 genotype and drug resistant mutation site detection kit and application thereof. The kit comprises set of primers detecting to-be-detected HIV-1 genome's genotype and drug resistant mutation site. The set of primers include amplification primer pairs and sequencing primer sets. Experiments prove that the detection kit provided by the invention ensures the detection capability of low-frequency mutant strains, greatly makes up the disadvantages of poor amplification efficiency, low detection rate and low low-frequency mutation detection rate of similar kits imported from abroad to China HIV epidemic strains, has the characteristics of simple and fast operation, high efficiency, economical efficiency and high throughput, etc., not only lays a solid foundation for domestic production of HIV drug resistant detection reagents, but also opens up broad prospect for domestic large-scale detection of HIV virus strain drug resistant situation.

Description

technical field [0001] The invention relates to a detection kit for HIV-1 genotype and drug resistance mutation site and application thereof, belonging to the field of biotechnology. Background technique [0002] Since 1981 when the United States first discovered that AIDS (AIDS) was caused by human immunodeficiency virus (HIV) infection, AIDS has spread rapidly around the world. According to the report on the global AIDS epidemic jointly released by UNAIDS and the World Health Organization, as of 2009, the total number of infected people in the world was 33.4 million, the number of newly infected people was about 2.7 million, and the death toll was 2 million. , 2.1 million infected people under the age of 15, with more than 7,400 new infections every day worldwide. However, the number of HIV infections in my country is increasing at a rate of 3% every year, and the trend of epidemic development is severe. According to the estimated report on the AIDS epidemic in China rel...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/703C12Q1/6869C12Q2600/16
Inventor 饶健郭弘妍田婕杨静静尚红韩晓旭赵彬张旻邢婉丽程京
Owner CAPITALBIO CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap