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Development of C-TILDA: A modified TILDA method for reservoir quantification in long term treated patients infected with subtype C HIV-1.
Journal of Virological Methods ( IF 2.2 ) Pub Date : 2019-11-19 , DOI: 10.1016/j.jviromet.2019.113778
Alessia Bertoldi 1 , Vanessa D'Urbano 1 , Isabella Bon 1 , Annelies Verbon 2 , Casper Rokx 2 , Charles Boucher 3 , Jeroen J A van Kampen 3 , Rob A Gruters 3 , Giorgio Gallinella 4 , Leonardo Calza 5 , Tokameh Mahmoudi 6 , Elisa De Crignis 6 , Maria Carla Re 1
Affiliation  

A better characterization of the HIV reservoir is pivotal for the development of effective eradication strategies. Accurate quantification of the latent reservoir remains challenging. Starting from a regular blood draw, the Tat/Rev induced limiting dilution assay (TILDA) combines serial dilution of CD4+ T cells with a PCR-based detection of HIV-1 spliced mRNA produced upon cell stimulation. Here we adapted the original protocol for HIV-1 subtype B to detect tat/rev mRNAs transcribed from reactivated latently infected cells in long term suppressed patients infected with HIV-1 subtype C. Given the heterogeneity of global HIV epidemiology, it is pivotal to develop assays with optimal performances also in patients infected with non-B subtypes. We observed that, in these patients infected with subtype C virus, the HIV reservoir quantified by TILDA correlates with both the time of virological suppression and CD4/CD8 ratio.

中文翻译:

C-TILDA的发展:一种改良的TILDA方法,用于对长期感染C型HIV-1的患者进行储库量化。

更好地表征艾滋病毒库对于制定有效的消灭策略至关重要。潜在储层的准确定量仍然具有挑战性。从定期抽血开始,Tat / Rev诱导的有限稀释法(TILDA)将CD4 + T细胞的连续稀释与细胞刺激后产生的HIV-1剪接的mRNA的基于PCR的检测结合起来。在这里,我们调整了HIV-1亚型B的原始协议,以检测在长期抑制的感染HIV-1亚型C的患者中从激活的潜伏感染细胞转录的tat / rev mRNA。鉴于全球HIV流行病学的异质性,开发至关重要在非B亚型感染的患者中也可实现最佳性能的检测。我们观察到,在这些感染了C型亚型病毒的患者中,
更新日期:2019-11-01
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