Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
Department of Dermatology and Venereology, All India Institute of Medical Sciences, New Delhi, India.
Mol Biotechnol. 2023 Dec;65(12):2099-2107. doi: 10.1007/s12033-023-00688-0. Epub 2023 Mar 24.
Neisseria gonorrhoeae (NG) is the second most common bacterial sexually transmitted infection (STI) worldwide. Gonorrhoea is a very serious infection because if untreated, it can lead to significant ramifications to reproductive, maternal, & newborn health and increase transmission of HIV. Infections are very often asymptomatic and symptoms when present manifest differently in men and women. The cornerstone of gonorrhoea control is to assure rapid diagnosis and prompt treatment of patients to prevent the onward spread of infection. The resource-rich settings are utilizing nucleic acid amplification tests (NAATs) for diagnosis, whereas resource-limited settings like ours where laboratory infrastructure is lacking, reliance is placed on syndromic approach. In view of the limitations of each, there is a compelling need for development of a point of care test (POCT). Aptamers offer such potential. These are short oligonucleotides that bind to its target with high affinity and specificity and therefore can be maneuvred for use in diagnostics. In this study, we performed live cell-SELEX (Systematic Evolution of Ligands by EXponential enrichment) to select 12 single-stranded DNA (ssDNA) aptamers that bind strongly to a cocktail of Neisseria gonorrhoeae strains, with Kd values ranging from 8.58 to 596 nM. Gold nanoparticle (GNP) assay revealed that one of the aptamers, E8 19 was highly specific for Neisseria gonorrhoeae (Kd = 24.5 nM). More importantly, it did not demonstrate any binding to Neisseria meningitidis and commensal Neisseria sp. The identified aptamer holds much promise for the development of a rapid test for diagnosis of gonorrhoea.
淋病奈瑟菌（NG）是全球第二常见的细菌性性传播感染（STI）。淋病是一种非常严重的感染，如果不治疗，可能会对生殖、产妇和新生儿健康产生重大影响，并增加 HIV 的传播。感染通常无症状，出现症状时，男性和女性的表现也不同。淋病控制的基石是确保迅速诊断和及时治疗患者，以防止感染的进一步传播。资源丰富的环境利用核酸扩增试验（NAAT）进行诊断，而像我们这样的资源有限的环境缺乏实验室基础设施，依赖于综合征方法。鉴于每种方法的局限性，迫切需要开发一种即时护理测试（POCT）。适体提供了这种潜力。这些是与靶标具有高亲和力和特异性结合的短寡核苷酸，因此可以用于诊断。在这项研究中，我们进行了活细胞 SELEX（通过指数富集的配体系统进化），以选择 12 种与淋病奈瑟菌菌株混合物强烈结合的单链 DNA（ssDNA）适体，其 KD 值范围为 8.58 至 596 nM。金纳米颗粒（GNP）测定表明，适体之一 E8 19 对淋病奈瑟菌具有高度特异性（KD = 24.5 nM）。更重要的是，它与脑膜炎奈瑟菌和共生奈瑟菌没有任何结合。鉴定出的适体为开发快速诊断淋病的测试提供了很大的希望。
