The ViroSeq HIV-1 Genotyping System, from Abbott GmbH, is a fully capillary- based genetic analysers (ABI PRISM , Avant, , , and Natalia M Marlowe at Abbott Laboratories The new Applied Biosystems ViroSeq HIV-1 Genotyping System (HGS) was formally released in. In this study, the Abbott RealTime HIV-1 (Abbott RealTime) assay was compared to the Roche Cobas TaqMan HIV-1 (Cobas TaqMan) and the.
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Genotypic analysis of HIV-1 drug resistance mutations | Scientist Live
Sven Thamm looks at a fully integrated and complete solution for detecting and reporting HIV-1 resistance to specific anti-retroviral therapy. Drug treatment of HIV infected patients includes use of multiple antiretroviral drugs in a strategy known as anti-retroviral therapy ART. ART usually includes adherence of three different drugs at several time points per day. Currently, anti-retroviral therapeutics are used for inhibiting the activity of the protease and reverse hic RT and thereby inhibiting the replication of the virus.
However, during the replication of the virus the reverse transcriptase often makes mistakes that may abbott to mutations that cause resistance against the drug.
This is often the cause of viral rebound and failure of the therapy. Identification vifoseq the presence of patient-specific drug resistant virus populations by genotypic resistance tests, can help the clinician to select an effective combination of drugs that virosfq likely to suppress HIV-1 replication sufficiently. These guidelines also suggest genotyping of infected mothers and their newborns.
ViroSeq detects mutations in the RT and protease genes of the pol region and provides the physician with a report indicating genetic evidence of viral resistance to known ART.
The Abbltt HIV-1 Genotyping System consists of different modules for sample preparation, reverse transcription, polymerase chain reaction RT-PCRcycle sequencing, automated electrophoresis and detection and analysis, and results interpretation.
The variation in throughput serves diagnostic laboratories in low, medium, and high throughput segments.
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The virus particles are isolated by centrifugation of the patient’s plasma. This is followed by a lysis and precipitation to isolate the RNA.
In the one-tube amplification steps a 1. A built in contamination control protects carryover from other samples by using Uracil-N-glycosylase. Due to the incorporation of dUTPs into the PCR product, cross contaminations from previously amplified patient samples will be digested by Uracil-N-glycosylase.
This digestion is initiated before the first viroeeq of the PCR. The PCR product is finally purified and checked for yield on a 1 per cent agarose gel.
Genotypic analysis of HIV-1 drug resistance mutations
This approach enables a one-tube sequencing reaction and in contrast to other methods there is no need to split samples into four different reactions. By using this seven primers approach, the vast majority of the generated sequence is available as double stranded at minimum. Cycle sequencing products are cleaned up by precipitation or by column purification in order to remove the unincorporated BigDye Terminators. After denaturation samples are placed into the instrument to start electrophoresis.
For using the sequencer samples have to be loaded hiiv on the gel. The ViroSeq HIV-1 Genotyping System software combines the sequence data obtained hov the seven primers of each patient sample into a single project Fig.
The vast majority of the sequence information is available from both strands for optimal reliability of the sequence. II A text mutation report showing a list of mutations found and categorised as known or unknown. Read the latest newsbrief.
Skip to main content. Drug identification Identification of the presence of patient-specific drug resistant virus populations by genotypic resistance tests, can help the clinician to select an effective combination of drugs that are likely to suppress HIV-1 replication sufficiently. New research looks at novel ways to combat drug resistance.
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