Method for detecting activity of antivirus medicament and application thereof

An antiviral drug, activity detection technology, applied in biochemical equipment and methods, microbial determination/inspection, material thermal development, etc.

Inactive Publication Date: 2011-02-02
302 MILITARY HOSPITAL OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not every virus can produce cytopathic changes in cell culture, and the judgment of CPE is highly subjective and has poor accuracy, so it is not suitable for accurate calculation of virus infectivity and judgment of drug antiviral activity

Method used

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  • Method for detecting activity of antivirus medicament and application thereof
  • Method for detecting activity of antivirus medicament and application thereof
  • Method for detecting activity of antivirus medicament and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Implementation method:

[0048] (1), select the mumps virus corresponding to Radix Radix isatidis heat-clearing and detoxifying activity indication;

[0049] (2), virus transfection Hela cells: take 37 ℃, 5% CO 2 Hela cells after 24h culture, add 100TCID 50 The mumps virus solution was used to transfect the Hela cells, and then the virus solution was discarded;

[0050] (3) Preparation of medicinal materials and chemical parts: Radix isatidis. Take 500g of Radix Radix Radix, decoct in water for 2 times, combine the decoction and concentrate into extract to obtain the total extract of the decoction (Part I, 156.6g), and extract with different polar solvents to obtain petroleum ether extract (Part II, 0.19g), ethyl acetate extract (Part III, 0.49g), chloroform extract (Part IV, 0.55g), n-butanol extract (Part V, 5.12g) and extracted substance (Part VI Parts, 116.25g) and other parts of different polarities. In the in vitro cell test, the II-V extraction parts were...

Embodiment 2

[0065] The steps of this example are the same as those of Example 1, the difference is that before the step (1) of Example 1, different extraction parts of Radix Radix are carried out at 0-8mg / ml -1 Perform cytotoxicity analysis on Hela cells at 8 concentrations within the range, the specific steps are:

[0066] Dilute the cell suspension to 1×10 6 cells / ml, seeded in a 96-well plate, 100 μL per well, placed at 37°C, 5% CO 2 After culturing in the incubator for 24 hours, add the test solution of parts I to VI according to the gradient dilution respectively, and the drug concentration is 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.063mg / ml -1 , continue to cultivate under the same conditions for 48h. Repeat 4 wells for each drug concentration, and set up normal cell controls in addition to observe the cytopathic changes (CPE) situation, the cytopathic changes are completely destroyed as 4; 75% is 3; 50% is 2; no pathological changes is 0 . Calculate the average cytopathic degree and inh...

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Abstract

The invention discloses a method for detecting activity of an antivirus medicament, and belongs to the technical field of the method for detecting property of the medicament. The method comprises the following steps of: (1) selecting microorganisms or cells corresponding to treatment indication of a to-be-detected medicament; (2) adding the to-be-detected medicament into the microorganisms or cells selected in the step (1) for culturing; and (3) placing the microorganisms or cells in which the to-be-detected medicament is added into a trace calorimeter to obtain a thermo-gram curve for growth and metabolism of the microorganisms or cells, and measuring the activity of the medicament according to the slope of the thermo-gram curve and the maximum heating power. The invention also comprises application of the detection method to measuring the heat clearing and detoxication activity of indigowoad root. The method of the invention can represent characteristic information during intervention of the microorganisms by the medicament, can acquire real-time online dynamic data and has the characteristics of real-time, online, high speed and accuracy.

Description

technical field [0001] The invention belongs to the technical field of drug property detection methods, and in particular relates to the establishment and application of an antiviral drug activity detection method. Background technique [0002] With the development of the medical field, people pay more and more attention to the effective parts of drugs and the application of these effective parts in the treatment of diseases in order to safely use drugs to avoid drug poisoning. At present, the detection method of the effective part of the drug and the dosage of the drug for the treatment of a certain disease usually adopts the method of first cultivating virus microorganisms or virus cells to make a certain macroscopic appearance on the surface of the visible carrier such as the culture medium, and then passing the drug to be tested through Physical and chemical means are added to the living environment of virus microorganisms or virus cells, thereby preventing the reproduct...

Claims

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Application Information

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IPC IPC(8): G01N25/20C12Q1/02
Inventor 肖小河赵艳玲王伽伯张萍
Owner 302 MILITARY HOSPITAL OF CHINA
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