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用于检测HIV蛋白酶的高亲和力适配体的开发与表征。

Development and characterization of high-affinity aptamers for HIV protease detection.

作者信息

Valadés-Alcaraz Ana, Reinosa Roberto, González-Hevilla Mario, Medina-Sánchez Carlos, Holguín África

机构信息

HIV-1 Molecular Epidemiology Laboratory, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Microbiology Department, Hospital Universitario Ramón y Cajal, CIBER en Epidemiología y Salud Pública (CIBERESP), Madrid, 28034, Spain.

出版信息

Heliyon. 2024 Sep 20;10(22):e38234. doi: 10.1016/j.heliyon.2024.e38234. eCollection 2024 Nov 30.

DOI:10.1016/j.heliyon.2024.e38234
PMID:39624307
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11609250/
Abstract

BACKGROUND

There are 39 million people infected by the human immunodeficiency virus (HIV) and 1.3 million new annually infections with more than 150 variants circulating. Early HIV detection is crucial for timely antiretroviral therapy and transmission prevention, but no technique can detect HIV before 10 days of infection. Aptamers, single-stranded oligonucleotides, can recognize specific proteins with high affinity and specificity. New molecular assays based on aptamers selected against highly-conserved peptides in HIV proteins could help to detect the virus earlier.

RESULTS

We have selected and characterized the first two aptamers against a highly-conserved and exposed peptide in the HIV protease (PR) after analyzing 236,240/6,436 HIV-1/HIV-2 PR sequences of nearly 150 HIV variants (types/groups/subtypes/circulating recombinants) by new bioinformatics tools. The selected aptamers were characterized by bioinformatics analysis, exploring their affinity, sensitivity, specificity, and best combination by Enzyme-Linked-Oligonucleotide-Assays (ELONA). Different strategies were explored to increase sensitivity for HIV PR detection, and the best one could detect viral PR (3.13 ng, 3.76·10 virions/well) and also HIV in infected samples with viraemia >8 log.

SIGNIFICANCE

Our anti-HIV PR aptamers are a promissing tool to be incorporated into new nanotechnologies for an ultrasensitive detection of HIV proteins in clinical samples for an early diagnosis of all virus variants.

摘要

背景

全球有3900万人感染了人类免疫缺陷病毒（HIV），每年新增感染病例130万，有超过150种病毒变体在传播。早期检测HIV对于及时进行抗逆转录病毒治疗和预防传播至关重要，但目前尚无技术能在感染10天前检测到HIV。适体是单链寡核苷酸，能以高亲和力和特异性识别特定蛋白质。基于针对HIV蛋白中高度保守肽段筛选出的适体的新型分子检测方法，可能有助于更早地检测该病毒。

结果

我们通过新的生物信息学工具分析了近150种HIV变体（类型/组/亚型/循环重组体）的236240/6436条HIV-1/HIV-2蛋白酶（PR）序列后，筛选并鉴定了首批两种针对HIV蛋白酶中高度保守且暴露的肽段的适体。通过生物信息学分析对所选适体进行了表征，利用酶联寡核苷酸分析（ELONA）探究了它们的亲和力、敏感性、特异性及最佳组合。探索了不同策略以提高对HIV PR检测的敏感性，最佳策略能够检测到病毒PR（3.13 ng，3.76·10个病毒体/孔），还能检测到病毒血症>8 log的感染样本中的HIV。

意义

我们的抗HIV PR适体是一种很有前景的工具，可纳入新的纳米技术中，用于超灵敏检测临床样本中的HIV蛋白，以早期诊断所有病毒变体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/6a20c4c62335/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/5ca5ea63b872/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/d815dc8a5524/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/73109c0f38e4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/82d774b334e2/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/6a9534bc347b/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/e57c7ab164e8/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/6a20c4c62335/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/5ca5ea63b872/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/d815dc8a5524/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/73109c0f38e4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/82d774b334e2/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/6a9534bc347b/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/e57c7ab164e8/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a6d/11609250/6a20c4c62335/gr6.jpg

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