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Generating aptamers towards human sperm cells using massively parallel sequencing.

James Gooch1, Sireethorn Tungsirisurp1, Hayley Costanzo1

  • 1Department of Analytical, Environmental & Forensic Sciences, King's College London, London, SE1 9NH, UK.

Analytical and Bioanalytical Chemistry
|August 6, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed novel DNA aptamers for accurate sperm cell detection, improving sexual offense investigations. These aptamers offer a sensitive and specific alternative to traditional methods for forensic analysis.

Keywords:
AptamersMassively parallel sequencingMicroscale thermophoresisNext-generation sequencingSELEXSperm cells

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Area of Science:

  • Forensic Science
  • Biotechnology
  • Molecular Biology

Background:

  • Sperm cell detection is vital in sexual offense investigations.
  • Traditional histological staining lacks specificity and sensitivity, complicating analysis.
  • Aptamer-based detection offers a promising alternative for improved sperm identification.

Purpose of the Study:

  • To select and characterize single-stranded DNA (ssDNA) aptamers that specifically bind to human sperm cells.
  • To develop a more sensitive and specific method for sperm detection compared to existing techniques.
  • To evaluate the potential of these aptamers in enhancing sexual offense casework.

Main Methods:

  • Utilized Cell-SELEX (Systematic Evolution of Ligands by EXponential enrichment) over 14 rounds with modified protocols for sperm isolation.
  • Employed massively parallel sequencing on the Illumina Miseq platform to identify potential aptamer sequences.
  • Developed a custom bioinformatics pipeline using Galaxy for automated analysis of sequencing data.
  • Validated aptamer binding specificity and affinity using microscale thermophoresis and enzyme-linked oligonucleotide assays.

Main Results:

  • Identified several promising ssDNA aptamer candidates with selective binding to human sperm cells.
  • Demonstrated high specificity and sensitivity of the selected aptamers in binding assays.
  • The developed bioinformatics pipeline efficiently processed large sequencing datasets.

Conclusions:

  • Selected ssDNA aptamers show significant potential for precise and efficient sperm cell detection.
  • These aptamers can overcome limitations of traditional methods, improving forensic analysis.
  • The developed aptamer-based approach could enhance the efficiency of sexual offense investigations.