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. 2019 Mar 13;14(3):e0212562.
doi: 10.1371/journal.pone.0212562. eCollection 2019.

Automated smartphone-based system for measuring sperm viability, DNA fragmentation, and hyaluronic binding assay score

Irene Dimitriadis  1 , Charles L Bormann  1 , Manoj Kumar Kanakasabapathy  2 , Prudhvi Thirumalaraju  2 , Hemanth Kandula  2 , Vinish Yogesh  2 , Neeraj Gudipati  2 , Vignesh Natarajan  2 , John C Petrozza  1 , Hadi Shafiee  2   3
Affiliations

Automated smartphone-based system for measuring sperm viability, DNA fragmentation, and hyaluronic binding assay score

Irene Dimitriadis et al. PLoS One. .

Abstract

The fundamental test for male infertility, semen analysis, is mostly a manually performed subjective and time-consuming process and the use of automated systems has been cost prohibitive. We have previously developed an inexpensive smartphone-based system for at-home male infertility screening through automatic and rapid measurement of sperm concentration and motility. Here, we assessed the feasibility of using a similar smartphone-based system for laboratory use in measuring: a) Hyaluronan Binding Assay (HBA) score, a quantitative score describing the sperm maturity and fertilization potential in a semen sample, b) sperm viability, which assesses sperm membrane integrity, and c) sperm DNA fragmentation that assesses the degree of DNA damage. There was good correlation between the manual analysis and smartphone-based analysis for the HBA score when the device was tested with 31 fresh, unprocessed human semen samples. The smartphone-based approach performed with an accuracy of 87% in sperm classification when the HBA score was set at manufacturer's threshold of 80. Similarly, the sperm viability and DNA fragmentation tests were also shown to be compatible with the smartphone-based system when tested with 102 and 47 human semen samples, respectively.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Smartphone-based semen analysis system.
(A) The actual smartphone optical attachment along with a smartphone. (B) The exploded image shows the various components of the smartphone-based semen analysis system. (C) Images acquired with the smartphone imaging platform for the three different assays, (i) HBA, (ii) viability, and (iii) DNA fragmentation.
Fig 2
Fig 2. Comparison of conventional method of analysis against automated smartphone-based HBA assessment.
(A) A linear regression analysis revealed strong agreement between the two methods (n = 31). The regression line is the solid line and the equation presented, is the line equation. (B) The samples were also compared using the Bland-Altman method of analysis (n = 31). The analysis revealed an absence of proportional and systematic bias for the tested sample set. The solid red line marks the mean bias. The dotted lines are the 95% limits of agreement of the sample set. (C) The scatter plot here represents the device performance in classifying samples as positive (<80%) and negative (= >80%). The system showed a sensitivity of 100% and a specificity of 69.23% (n = 31). The overall accuracy of the system in HBA-score based classification was 87.10% (D). The scatter plot shows the concentration and motility values of semen samples as measured by a CASA system along with its respective HBA score (n = 30).
Fig 3
Fig 3. Comparison of conventional method of analysis against using smartphone-based optics to measure sperm viability.
(A) Passing-Bablok analysis revealed a strong linear relationship between the smartphone-based analysis and conventional method of sperm viability assessments (n = 102). The regression line is the solid blue line and the equation presented, is the line equation. The red lines represent the 95% confidence intervals of the regression line. The black line represents the line of identity, (B) The Bland-Altman method of analysis (n = 102) was performed on the sample set. The blue line represents the mean bias. The red lines are the 95% limits of agreement of the sample set.
Fig 4
Fig 4. Comparison of conventional method of analysis against automated smartphone-based sperm DNA fragmentation assessment.
(A) The Bland-Altman method of analysis (n = 20) was performed on the sample set comparing the DNA fragmentation scores obtained through conventional analysis and the scores obtained manually through the use of a smartphone-based approach. (B) The Bland-Altman method of analysis (n = 27) was performed to compare the smartphone-based automated approach and the smartphone-based manual approach. The blue lines represent the mean bias while the red lines are the 95% limits of agreement of the sample set.
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References

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