Path and method for monitoring non nucleotide reverse transcriptive enzyme inhibitor anti retrovirus therapy

A technology of anti-retrovirus and reverse transcriptase inhibition, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, recombinant DNA technology, etc., and can solve problems such as inappropriate sensitivity, reduced resistance, mutation, etc.

Inactive Publication Date: 2004-09-29
病毒科学公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This suggests that the codon 181 mutation is not suitable for in vivo screening since it can abolish zidovudine resistance in vitro allowing reduced sensitivity to this NNRTI drug with concomitant zidovudine resistance mutations,

Method used

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  • Path and method for monitoring non nucleotide reverse transcriptive enzyme inhibitor anti retrovirus therapy
  • Path and method for monitoring non nucleotide reverse transcriptive enzyme inhibitor anti retrovirus therapy
  • Path and method for monitoring non nucleotide reverse transcriptive enzyme inhibitor anti retrovirus therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Determination of drug phenotypic sensitivity and resistance using resistance test vehicle

[0133] The methods and methods described in the PCT International Application (Application No. PCT / US / 01609, application date January 29, 1997) were used to conduct drug phenotypic sensitivity and resistance tests.

[0134] There is a segment from the patient (PDS) in these trials. This fragment is either a fragment of viral RNA isolated from viral particles in the serum of HIV patients, and then amplified from viral RNA by reverse transcription-polymerase chain reaction; or a paternal clone of resistance test vector DNA Site-directed mutagenesis was performed to obtain a wild-type HIV-1 mutant. This segment from the patient corresponds to the HIV protease and reverse transcriptase regions. Viral RNA is isolated using standard operating procedures, such as RNAgents Total RNA Isolation System (Promagnet, Madison, Wisconsin, USA) or RNAzol (Tail-Tester, Flanders, Texas, USA). The rever...

Embodiment 2

[0143] Example 2: Analyze genotype to establish the correlation between genotype and phenotype sensitivity

[0144] Phenotypic sensitivity analysis of patients' HIV samples

[0145] The resistance test vector was constructed as described in Example 1. Resistance test vectors or clones from the resistance test vector library are subjected to phenotypic sensitivity tests to accurately and quantitatively determine the sensitivity level of a series of antiretroviral drugs. This group of antiretroviral drugs may include the so-called nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleotide reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PRIs). If new drugs or new drug targets are available, this group of drugs can be expanded. On each resistant vector library, determine the IC50 of each test drug. Compare the sensitivity models of all tested drugs with known sensitivity models. Further examination of patient samples can be performed to identify genotypic c...

Embodiment 3

[0151] Establish the correlation between P225H genotype analysis and phenotypic sensitivity

[0152] Phenotype analysis of patients 97-302

[0153] The resistance test vector was constructed using patient 97-302 samples as described in Example 1. The patient has been treated with d4T, indinavir and DMP-226 for approximately 10 months. Isolate the viral RNA and perform reverse transcription-PCR to amplify a fragment from the patient. This fragment includes a viral fragment encoding the full length of the protease and reverse transcriptase 1-313 amino acids. The fragment was inserted into a viral vector with a reporter gene, and the resistance test vector was named RTV-302. Use phenotypic sensitivity test to accurately and quantitatively determine the sensitivity level of RTV-302 to a series of antiretroviral drugs. These drugs include the said NRTI drugs (AZT, 3TC, d4T, ddI, ddC), NNRTIs (delafurdine, nefurapine) and PRIs (indinavir, nelfinavir, ritonavir, saquinavir). Determine th...

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Abstract

This invention relates to antiviral drug susceptibility and resistance tests to be used in identifying effective drug regimens for the treatment of human immunodificiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) and further relates to the means and methods of monitoring the clinical progression of HIV infection and its response to antiretroviral therapy, particularly non-nucleoside reverse transcriptase inhibitor therapy using phenotypic susceptibility assays or genotypic assays.

Description

[0001] This application is a divisional application of the Chinese patent application number 99808689.4, the international filing date is May 26, 1999, and the invention title is "Methods and Methods for Monitoring Non-nucleoside Reverse Transcriptase Inhibitor Antiretroviral Therapy". [0002] This application is a partial continuation application of U.S. Patent Application No. 09 / 085,148 filed on May 26, 1998, and claims the rights and interests of U.S. Provisional Application No. 60 / 124,090 filed on March 12, 1999, the contents of which are incorporated herein as reference. [0003] In this application, various references are indicated in brackets. The entire contents disclosed in these documents are incorporated herein by reference to more fully describe the state of the art in the technical field to which this application belongs. Technical field [0004] The invention relates to an antiretroviral drug sensitivity and resistance test, which is used to identify effective drug p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/68C12Q1/70
CPCC12Q2600/156C12Q2600/106C12Q1/703
Inventor 珍尼特·惠特科姆
Owner 病毒科学公司
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