Department of Analytical, Environmental & Forensic Sciences, King's College London, London, SE1 9NH, UK.
School of Life Sciences, The University of Warwick, Coventry, CV4 7AL, UK.
Anal Bioanal Chem. 2021 Sep;413(23):5821-5834. doi: 10.1007/s00216-021-03562-7. Epub 2021 Aug 5.
Determining the presence of sperm cells on an item or swab is often a crucial component of sexual offence investigation. However, traditional histological staining techniques used for the morphological identification of spermatozoa lack both specificity and sensitivity, making analysis a complex and time-consuming process. New methods for the detection of sperm cells based on aptamer recognition may be able to overcome these issues. In this work, we present the selection of ssDNA aptamers against human sperm cells using Cell-SELEX and massively parallel sequencing technologies. A total of 14 rounds of selection were performed following a modified Cell-SELEX protocol, which included additional steps for the isolation of spermatozoa from seminal fluid. Massively parallel sequencing using the Illumina Miseq platform was conducted on enriched aptamer pools to elucidate the structure of potential binders. A custom bioinformatics pipeline was also developed using Galaxy for the automated processing of sequencing datasets. This data revealed several promising aptamer candidates, which were shown to selectively bind sperm cells through both microscale thermophoresis and enzyme-linked oligonucleotide assays. These aptamers have the potential to increase the efficiency of sexual offence casework by facilitating sperm detection.
确定物品或拭子上是否存在精子细胞通常是性犯罪调查的关键组成部分。然而，传统的用于鉴定精子形态的组织学染色技术既缺乏特异性又缺乏敏感性，使得分析成为一个复杂且耗时的过程。基于适配体识别的新方法可能能够克服这些问题。在这项工作中，我们展示了使用 Cell-SELEX 和大规模平行测序技术对人精子细胞进行 ssDNA 适配体选择的方法。在经过修改的 Cell-SELEX 方案的指导下，总共进行了 14 轮选择，该方案包括从精液中分离精子的额外步骤。使用 Illumina Miseq 平台对富集的适配体池进行大规模平行测序，以阐明潜在结合物的结构。还使用 Galaxy 开发了一个自定义的生物信息学管道，用于自动处理测序数据集。该数据揭示了一些有前途的适配体候选物，这些候选物通过微尺度热泳和酶联寡核苷酸测定被证明能够选择性地结合精子细胞。这些适配体有可能通过促进精子检测来提高性犯罪案件工作的效率。
