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US 2018/0003666 A1Published application
Highly sensitive method for detection of viral hiv dna remaining after antiretroviral therapy of aids patients
OwnerLuc Montagnierinventor
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Abstract
Methods for detecting polynucleotides, especially the DNA replicated from samples obtained from subjects infected with pathogenic viruses such as human immunodeficiency virus, by detecting electromagnetic signals (“EMS”) emitted by such polynucleotides, and methods for improving the sensitivity of the polymerase chain reaction (“PCR”).
Claims20
A method for detecting a polynucleotide comprising:
- obtaining an clinical specimen from a human;
- isolating deoxyribonucleic acids from the clinical specimen;
- performing a polymerase chain reaction amplification on the clinical specimen, using primers specific for at least one of gene sequence of HIV;
- and
- detecting a 104 bp amplicon having no more than a 2 bp difference from a 104 bp portion of HIV DNA LTR.
2 more elements
A method for detecting a polynucleotide comprising:
- obtaining a specimen from a living organism;
- isolating deoxyribonucleic acids from the specimen;
- treating the isolated deoxyribonucleic acids from the specimen with RNase, to degrade RNA present in the isolated deoxyribonucleic acids;
- after treatment with RNase, treating the isolated deoxyribonucleic acids with reverse transcriptase;
- after treatment with RNase, performing a polymerase chain reaction amplification on the clinical specimen, using primers specific for at least one pathogen of the living organism;
- and
- selectively detecting an amplicon associated with the primers,
- wherein a sensitivity of detection of the amplicon from the polymerase chain reaction amplification of the isolating deoxyribonucleic acids corresponding to the primers is increased by the treatment with reverse transcriptase, as compared to a sensitivity absent the reverse transcriptase treatment.
5 more elements
The method according to claim wherein the RNase and reverse transcriptase are distinct enzymes.
A method of detecting an HIV deoxyribonucleic acid, comprising:
- obtaining an clinical specimen from a human comprising washed red blood cells;
- isolating deoxyribonucleic acids from the clinical specimen;
- treating the isolated deoxyribonucleic acids from the clinical specimen with RNase, to degrade RNA present with the isolating deoxyribonucleic acids;
- treating the RNase-treated isolating deoxyribonucleic acids with a reverse transcriptase, using the isolating deoxyribonucleic acids as a template;
- performing a polymerase chain reaction amplification on the clinical specimen using Taq polymerase, using HIV gene-specific primers;
- and
- detecting an amplicon which is homologous with corresponding to a portion of the HIV DNA having a length of at least 104 bp with no more than a 2 bp difference within the at least 104 bp,
- wherein the reverse transcriptase treatment increases a sensitivity of the detection of the amplicon as compared to the absence of the reverse transcriptase treatment.
5 more elements
Dependencies are inferred from the claim text.