The Applied Biosystems HIV-1 Genotyping Kit with Integrase is designed to genotype the protease, reverse transcriptase (RT), and integrase (IN) regions of the HIV-1 pol gene. The genotyping result encompasses codons 6-99 in the PR region, codons 1-251 in the RT region, and codons 1-288 of the IN region of the gene. HIV-1 genomic mutations in these regions confer resistance to anti-retroviral drugs that act through the inhibition of the protease and reverse transcriptase.

The HIV-1 Genotyping Kit with Integrase enables reliable genotyping of the genetically diverse HIV-1 virus to investigate drug resistance mutations (DRMs) to protease inhibitors (PI), nucleoside reverse-transcriptase inhibitors (NRTI), non-nucleoside reverse-transcriptase inhibitors (NNRTI), and integrase inhibitors (INSTI) from HIV-1 RNA extracted from EDTA plasma and dried blood spot samples. This product employs assays for HIV-1 genotyping licensed from the U.S. Centers for Disease Control and Prevention.

The HIV-1 Genotyping Kit with Integrase contains amplification and sequencing reagents targeted to the protease, reverse transcriptase, and integrase regions of the HIV-1 pol gene.  Specifically, the kit contains ready-to-use master mix, enzyme, and positive and negative controls to cover the three phases of the workflow: RT-PCR, nested PCR, and bidirectional cycle sequencing steps to process 48 test samples.

Starting with viral RNA extracted from an EDTA plasma or dried blood spot sample, RT-PCR is conducted to generate PR/RT and IN PCR products as DNA template. After confirming the band size of the PCR amplicons, the Cycle Sequencing module is used to sequence the DNA templates utilizing Sanger sequencing technology. The sequencing reactions are first purified, then sequenced using POP-7 on a designated capillary electrophoresis system. The sequencing traces are reviewed for quality, after which the user exports the AB1 file for use in downstream analysis and reporting.

The HIV-1 Genotyping Kit with Integrase delivers gold-standard Sanger sequencing results for use in the identification of drug resistant mutations from the diverse and rapidly evolving HIV-1 virus.