Assessment of the feasibility of SP-10 as a male fertility marker and validation of a lateral flow immunoassay for indirect sperm concentration analysis

Mowla, Shahriar (2018) Assessment of the feasibility of SP-10 as a male fertility marker and validation of a lateral flow immunoassay for indirect sperm concentration analysis. MRes thesis, University of Nottingham.

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Over the past few decades, the emphasis on male gamete has increased as male factor infertility contributes to almost 30% of infertile couples. As a result, the need for development of more accurate assays is now more important than ever. Determination of the concentration of sperm using an indirect assay does not possess the inherent user subjectivity, procedure uncertainty and biological restrictions such as viscosity of manual semen analysis. This thesis aims to assess the feasibility of SP-10 protein to be used as a fertility marker by the assessment of a lateral flow immunoassay.

Preliminary analysis for sample preparation, handling and production outlined a suitable protocol for novel device analysis. Validation of a novel diagnostic sperm counting device as developed by Swiss Precision Diagnostics (SPD) against the laboratory semen analysis was the primary objective of the current study. Suitable concentration of lysis buffer, incubation period and device sensitivity and specificity were then determined by analysing patient samples seeking fertility advice.

A series of analysis results for the preliminary examinations, indicated that stirring the sample is an acceptable compromise to using a high speed vortex for homogenisation of semen. It was also found that the chosen lysis buffer at its current concentration is a suitable agent for acrosomal sperm membrane impairment for releasing SP-10 into the solution from the entire sperm population. Accuracy assessment of the device prototype showed that the accuracy of the device at the lowest threshold (5x106/ml)

expressed 100% accuracy compared with CASA and haemocytometer readings.

Further analysis is required to definitively accept the thresholds for the percentage absorbance of the high and low sensitivity strips inside the device; particularly at the 15x106/ml threshold where the device expressed 85% accuracy. The validation data shows that indirect measurement of seminal sperm concentration using the novel device can accurately estimate the sperm concentration. However, further optimisation is required before the device is ready for commercial distribution.

Item Type: Thesis (University of Nottingham only) (MRes)
Supervisors: Marsters, Peter
Tomlinon, Mathew
Keywords: Male infertility; Sperm count; Immunoassay; Fertility markers; Spermatozoa
Subjects: W Medicine and related subjects (NLM Classification) > WJ Urogenital system
Faculties/Schools: UK Campuses > Faculty of Medicine and Health Sciences > School of Medicine
Item ID: 49257
Depositing User: Mowla, Shahriar
Date Deposited: 16 Apr 2018 14:26
Last Modified: 17 Apr 2018 03:14

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