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Selection of DNA Aptamers Against Neisseria gonorrhoeae Causing Sexually Transmitted Infection (STI)

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Abstract

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.

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Acknowledgements

The work was supported by funding from Department of Health Research, Ministry of Health & Family Welfare, India (Grant No. DHR/HRD/WS-03-2015-16). The authors gratefully acknowledge Professor Monica M Lahra & Dr. Athena Limnios, Neisseria Reference Laboratory and WHO Collaborating Center for STD, SEALS Microbiology, The Prince of Wales Hospital, Australia for presenting commensal Neisseria sp. to our laboratory. We are also thankful to Dr. Benu Dhawan for providing Mycoplasma hominis NCTC 10111 and Ureaplasma urealyticum NCTC 10177. The authors acknowledge Shri Rajendra Singh for his technical assistance.

Funding

The work was supported by funding from Department of Health Research, Ministry of Health & Family Welfare, India (Grant No. DHR/HRD/WS-03–2015-16).

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Authors and Affiliations

  1. Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India

    Shilpi Malhotra & Seema Sood

  2. Department of Dermatology and Venereology, All India Institute of Medical Sciences, New Delhi, India

    Somesh Gupta

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  1. Shilpi Malhotra
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All authors contributed to the study conception and design. SM: Carried out the research work. The first draft of the manuscript was written by SM and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Seema Sood.

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Malhotra, S., Gupta, S. & Sood, S. Selection of DNA Aptamers Against Neisseria gonorrhoeae Causing Sexually Transmitted Infection (STI). Mol Biotechnol 65, 2099–2107 (2023). https://doi.org/10.1007/s12033-023-00688-0

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