Method for screening anti-aids drug

A screening method and anti-AIDS technology, applied in the field of molecular biology, can solve problems such as cost constraints and achieve accurate drug effects

Inactive Publication Date: 2013-05-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because HIV is infectious to humans, it must be strictly limited to negative pressure P3 laboratories, and the high cost restricts large-scale general screening of drugs

Method used

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  • Method for screening anti-aids drug
  • Method for screening anti-aids drug
  • Method for screening anti-aids drug

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, the selection of optimal cell ratio and optimal co-culture conditions

[0032] (1) Normal cell culture (that is, the cells can grow normally) JLTRG cells and H9 cells are cultured using 1640 medium containing 10% FBS, 1% streptomycin and 1% penicillin (that is, adding 100g per liter of 1640 medium FBS of 10g, streptomycin of 10g and penicillin of 10g), cell count; Generally, the number of suspension in every 2ml of 1640 culture medium containing 10% FBS, 1% streptomycin and 1% penicillin is 10 6 Cell. Cultured in a cell culture incubator at 37°C with CO 2 The concentration (volume concentration) is 5%, and the humidity (relative humidity) is 75%. The incubation time is generally 24 hours.

[0033] (2) Put JLTRG cells and H9 cells in 5% FBS, 1% streptomycin and 1% Penicillin 1640 medium co-cultured for 24 hours, 48 ​​hours, 72 hours, 96 hours. Cultured in a cell culture incubator at 37°C with CO 2 The concentration is 5% and the humidity is 75%. The nu...

Embodiment 2

[0039] Embodiment 2, take three kinds of existing medicines (T20, EFV, PTD) as experimental detection target, determine the practicability of the method of the present invention:

[0040] According to the experimental conditions screened above, JLTRG cells and H9 cells were co-cultured in 1640 medium with 5% FBS, 1% streptomycin and 1% penicillin at a cell ratio of 10:1.

[0041] (1) Dilute different drugs, T20 (0.00015uM, 0.0006uM, 0.3uM, 0.003uM, 0.015uM, 0.06uM),

[0042] PTD (1.5uM.0.75uM, 0.375uM, 0.18, 0.09, 0.045uM),

[0043] EFV (0.6uM, 0.15um, 0.03uM, 0.006uM, 0.0015, 0.0003uM);

[0044] Both use RPMI 1640 medium as the diluent.

[0045] (2) Cultivate the above-mentioned three types of drug solutions with different concentrations and JLTRG cells for 6 hours. The number of each hole is 10 6 The JLTRG cells were cultivated in a cell culture incubator at 37°C, and the CO in the cell culture incubator 2 The concentration is 5%, t...

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Abstract

The invention discloses a method for screening an anti-aids drug. The method comprises the following steps: 1) diluting the drug to be detected into drug solutions of different concentrations; 2) nurturing JLTRG cells in the drug solutions; 3) adding a 1640 culture medium containing 5% FBS, 1% streptomycin and 1% penicillin as well as H9 cells to perform co-culture; 4) measuring the expression condition of green fluorescence with a flow cytometer, and obtaining the corrected cell fluorescence value according to the percentage of the green fluorescence cells and the average fluorescence intensity; 5) separately setting positive control and negative control; and 6) measuring the effective dose resisting half of aids, of the drug to be measured according to the experiment result. The method disclosed by the invention can quickly and effectively implement screening of the anti-aids drug.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to an anti-AIDS drug screening technology and a research method. Background technique [0002] AIDS (AIDS) is a kind of human immunodeficiency virus (human immunodeficiency virus, referred to as HIV) that invades the human body and destroys the immune function of the human body, causing a variety of incurable infections and tumors in the human body, and finally leading to the death of the infected person. a serious infectious disease. Looking back over the past 25 years, the prevalence of AIDS has far exceeded people's fears and estimates. Until now, the medical community has not yet found the best solution to this disease. Relevant scientific research institutions are trying to find the weakness in the virus's own defense, so as to develop antiviral drugs targeting the virus itself and host factors in order to inhibit or kill the virus. the virus. [0003] The rapid rise...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/18C12R1/93
Inventor 王英杰吴南屏冯婷婷
Owner ZHEJIANG UNIV
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