title: Strategies to quantify unspliced and multiply spliced mRNA expression in HIV-2 infection.
abstract: HIV-2 infection is associated with a slower rate of disease progression with limited impact on the survival of the majority of infected adults, and much lower plasma viral load than HIV-1. In spite of the major differences in viremia, the quantitative assessment of HIV-2 proviral load documented levels similar to those observed in HIV-1 infected individuals, suggesting an equivalent number of circulating infected cells in both infections. It remains unclear whether this apparent paradox results from a contribution of latent/quiescent viruses or from transcriptional and/or post-transcriptional control of HIV-2 replication. In order to investigate these possibilities, a one-step and two-step reverse transcription quantitative real-time PCR based methods (RT-qPCR) for gag and tat mRNA HIV-2 transcripts were developed. These methods were validated and compared to assess the expression of HIV-2 gag and tat transcripts in parallel with proviral DNA and viral production. The results suggest that the two-step approach may allow a better detection of low level gag and tat mRNA HIV-2 transcripts.
authors.affiliation: Unidade de Imunologia Clínica, Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028 Lisboa, Portugal.
authors.forename: Rui S
authors.initials: RS
authors.lastname: Soares
authors.forename: Paula
authors.initials: P
authors.lastname: Matoso
authors.forename: Marta
authors.initials: M
authors.lastname: Calado
authors.forename: Ana E
authors.initials: AE
authors.lastname: Sousa
mesh_terms: D002478:Cells, Cultured; D015683:Gene Products, gag / Q000096:biosynthesis / Q000235:genetics; D015696:Gene Products, tat / Q000096:biosynthesis / Q000235:genetics; D015498:HIV-2 / Q000235:genetics*; D006801:Humans; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis; D012367:RNA, Viral / Q000032:analysis* / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014158:Transcription, Genetic
chemical_list: D015683:Gene Products, gag; D015696:Gene Products, tat; D012333:RNA, Messenger; D012367:RNA, Viral