Table 1.

Strengths and limitations of current commercially available ROS detection tools for male infertility.

Assay	Company	What it Measures	Strengths	Limitations
Luminol Chemiluminescence	Multiple companies	Luminol is first oxidized by many radicals (i.e., -OH and CO3) and peroxidases, forming the luminol radical. The luminol radical then reacts with superoxide, forming the short-lived intermediate hydroperoxide. Hydroperoxide is decomposed to 3-aminophyhalane, which emits light.	High quantum yield Highly sensitive Easy to measure Readily available Highly reproduceble	Cannot differentiate different ROS radicals Large semen volume required (~400 µL). Cannot differentiate mature sperm from immature sperm or other cell types (i.e., leucocytes). No defined references ranges/decision limits Cannot be used on frozen samples Temperature-sensitive >25 °C Light-sensitive and can oxidize over time
MiOXSYS	MiOXSYS	Measures the transfer of electrons from oxidants to antioxidants (sORP)	Rapid result (~5 min) Highly sensitive Small sample volume required (30 µL) Good reproducibility Obtain accurate results up to 2 h post ejaculation Can be used on fresh or frozen samples Cost effective	No definitive reference ranges/decision limits Cannot differentiate different ROS radicals Temperature-sensitive between 2 and 37 °C
OxiSperm	Halotech DNA	When exposed to superoxide, the NBT reagent is reduced and converted into blue formazan crystals.	Time-effective Highly sensitive Cost-effective Rapid result (~15 min) Small sample volume required (~30–50 µL)	Subjective analysis Low assay precision Only measures one ROS radical Seminal plasma reductase interference Not found to correlate with fertilization, embryo development or pregnancy outcomes in ART

ART: Assisted Reproductive Technology, Temp: Temperature.