Means and method for monitoring non-nucleoside reverse transcriptase inhibitor antiretroviral therapy

An anti-retroviral, reverse transcriptase technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, and microbial assay/inspection, etc.

Inactive Publication Date: 2001-09-05
病毒科学公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This suggests that the codon 181 mutation is not suitable for in vivo screening because it counteracts zidovudin...

Method used

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  • Means and method for monitoring non-nucleoside reverse transcriptase inhibitor antiretroviral therapy
  • Means and method for monitoring non-nucleoside reverse transcriptase inhibitor antiretroviral therapy
  • Means and method for monitoring non-nucleoside reverse transcriptase inhibitor antiretroviral therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Measuring Phenotypic Sensitivity and Resistance to Drugs Using Resistance Assay Vectors

[0132] Drug phenotype susceptibility and resistance tests were carried out using the means and methods described in PCT International Application (Application No. PCT / US / 01609, filing date: January 29, 1997).

[0133] One of these trials was a patient-derived segment (PDS). This fragment, either by isolating viral RNA from the virus particles in the serum of HIV patients, and then using reverse transcription-polymerase chain reaction to amplify a fragment from the viral RNA; or it is a paternal clone of the resistant test vector DNA Perform site-directed mutagenesis to obtain wild-type HIV-1 mutants. This patient-derived fragment corresponds to the HIV protease and reverse transcriptase regions. Viral RNA is isolated using standard operating procedures, such as the RNAgents Total RNA Isolation System (Promega, Madison, WI, USA) or RNAzol (Tyer-Test, Flanders Wood, TX, USA). The ...

Embodiment 2

[0142] Example 2: Analyzing Genotypes Establishing the Correlation Between Genotype and Phenotype Sensitivity

[0143] Phenotypic Sensitivity Analysis of Patient HIV Samples

[0144] Resistance test vectors were constructed as described in Example 1. Perform phenotypic susceptibility testing on resistance test vectors or clones from resistance test vector libraries to accurately and quantitatively determine susceptibility levels to a range 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). This group of drugs can be expanded if new drugs or new drug targets become available. On each resistant carrier library, the half inhibitory concentration IC50 of each test drug was determined. Sensitivity models for all test drugs were compared to known sensitivity models. Patient samples can be further examined...

Embodiment 3

[0150] Establishing the correlation between P225H genotype analysis and phenotypic sensitivity

[0151] Phenotype Analysis of Patient 97-302

[0152] The resistance test vector was constructed as described in Example 1 using patient 97-302 samples. The patient had been treated with d4T, indinavir, and DMP-226 for approximately 10 months. Viral RNA was isolated and subjected to reverse transcription-PCR to amplify a fragment from the patient. The fragment includes a viral fragment encoding the full length of the protease and 1-313 amino acids of the reverse transcriptase. The fragment was inserted into a viral vector with a reporter gene, and the resistance test vector was obtained and named RTV-302. The level of RTV-302 susceptibility to a range of antiretroviral drugs was accurately and quantitatively determined using a phenotypic susceptibility assay. These drugs include the said NRTI drugs (AZT, 3TC, d4T, ddI, ddC), NNRTIs (delavirdine, nevirapine) and PRIs (indinavir, ...

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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 continuation-in-part of U.S. Patent Application No. 09 / 085,148, filed May 26, 1998, and claims the benefit of U.S. Provisional Application No. 60 / 124,090, filed March 12, 1999, the contents of which are incorporated herein as refer to. [0002] Throughout this application various references are indicated in parentheses. The entire disclosures of these documents are incorporated herein by reference in order to more fully describe the state of the art in the technical field to which this application pertains. technical field [0003] The present invention relates to antiretroviral drug susceptibility and resistance assays for identifying effective pharmaceutical preparations for the treatment of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS). The invention further relates to monitoring the progression of clinical HIV infection and its response to antiretroviral therapy using phenotypic and genotypic assays...

Claims

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Application Information

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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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