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Primer composition for identifying HIV (human immunodeficiency virus) in assistance mode and application thereof

A primer composition and a technology for identifying people, which can be applied in the field of assisting HIV identification, and can solve the problems of blood safety threat, high risk of transmission, etc.

Active Publication Date: 2011-11-23
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although no HIV antibody can be detected during the window period, HIV already exists in the body, which is also contagious, and the risk of transmission is very high, especially posing a great threat to blood safety. Therefore, it is necessary to develop a test that can effectively detect infected persons during the window period approach is urgently needed to control the AIDS epidemic

Method used

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  • Primer composition for identifying HIV (human immunodeficiency virus) in assistance mode and application thereof
  • Primer composition for identifying HIV (human immunodeficiency virus) in assistance mode and application thereof
  • Primer composition for identifying HIV (human immunodeficiency virus) in assistance mode and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the design of primer composition

[0022] Take advantage of Vector NTI Advance TM 10 The analysis software scans the conserved regions of a total of 1582 protease region and reverse transcriptase region sequences (sequence composition as shown in Table 1) from HIV-infected persons, and observes the degree of variation of the pol region genes of different subtypes. The scanning results showed that the gene conservation of the protease region in the pol region was better than that of the reverse transcriptase region.

[0023] Table 1 Gene sequence composition of HIV-1 pol region

[0024]

[0025] For the results of scanning the conserved region of the protease region, see figure 1 . For the results of scanning the conserved region of the reverse transcriptase region, see figure 2 .

[0026] A group of primers (named as primer composition HIV-CN-II) are designed based on sequence analysis results, by primer HIV-CN-II-1, primer HIV-CN-II-2, primer HIV...

Embodiment 2

[0029] Embodiment 2, the sensitivity of primer composition

[0030] 1. Experimental samples

[0031] The plasma samples of 41 clinically confirmed HIV-infected persons (informed consent volunteers) were used as experimental samples, among which 17 were infected with HIV-1B' subtype strains, and 12 were infected with HIV-1CRF01-AE subtype 12 patients were infected with HIV-1B / C recombinant subtype strains.

[0032] 2. Application of primer composition

[0033] The plasma of each patient in step 1 was subjected to the following experiments:

[0034] 1. Extract total RNA.

[0035] 2, with the total RNA of step 1 as template, carry out RT-PCR (the first round of PCR amplification) with the primer pair that HIV-CN-II-1 and HIV-CN-II-2 form, obtain the first round of PCR amplification increase product.

[0036] The reaction system of the first round of PCR amplification is shown in Table 3.

[0037] Table 3 The reaction system of the first round of PCR amplification

[0038] ...

Embodiment 3

[0055] Embodiment 3, the specificity of primer composition

[0056] 1. Experimental samples

[0057] Five HTLV-infected patients who were clinically confirmed and positive for nucleic acid testing (volunteers who gave informed consent; numbers were HTLV25, HTLV26, HTLV36, HTLV37, HTLV38), and five HBV-infected patients who were clinically confirmed and positive for nucleic acid testing (informed consent volunteers). Volunteers; numbers are HBV-2, HBV-5, HBV-6, HBV-10, HBV-12), 7 HCV infected persons who were clinically confirmed and positive in nucleic acid test (informed consent volunteers; numbers are respectively HCV-5, HCV-71, HCV-75, HCV-76, HCV-93, HCV-7, HCV-56) as experimental samples.

[0058] 2. Application of primer composition

[0059] The plasma of each patient in Step 1 was subjected to the same experiment as Step 2 of Example 2.

[0060] The agarose gel of the second round of PCR amplification products is shown in Figure 5 . The amplified experimental samp...

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Abstract

The invention discloses a primer composition for identifying HIV (human immunodeficiency virus) in assistance mode and application thereof. The primer composition provided by the invention is composed of a DNA represented by a sequence 1 in a sequence table, a DNA represented by a sequence 2 in the sequence table, a DNA represented by a sequence 3 in the sequence table, and a DNA represented by a sequence 4 in the sequence table; the primer composition can be used for (a) identifying the human immunodeficiency virus (HIV) in assistance mode, or (b) identifying the HIV-infected patient in assistance mode. The primer composition has important application value in identification of HIV and diagnosis of HIV-infected patients in assistance mode, and has great influence on social health.

Description

technical field [0001] The invention relates to a method for assisting identification of HIV and its special primer composition. Background technique [0002] Human immunodeficiency virus (HIV) is the pathogen that causes human acquired immunodeficiency syndrome (AIDS). Since AIDS was first diagnosed in the United States in 1981, AIDS has spread rapidly around the world. AIDS has been prevalent in China for 25 years. As of the end of October 2010, China has reported more than 370,000 HIV-infected and AIDS patients, including more than 130,000 AIDS patients, and more than 68,000 deaths. After effectively controlling the transmission of paid donation / blood transfusion in the past, in recent years, with the development of society and economy and the effective deepening of AIDS prevention and control work, some new epidemic features have emerged in the AIDS epidemic: ①The AIDS epidemic is still rising, but the rate of increase is somewhat Slow down; ②Sexual transmission has bec...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/70C12Q1/68
Inventor 李韩平李敬云郭伟鲍作义李林刘永健庄道民刘思扬李天一王晓林
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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