Multiple Biomarkers To Improve The Diagnostic Accuracy

The study on the optimizing combination of multiple biomarkers; to improve the diagnostic accuracy in male fertility. However, only 50% of successful full term pregnancy rates per insemination have been reporting. Half of all breeding failures are due to male factor infertility or sub fertility; leading to a decrease in living offspring production and huge economic losses. Therefore, the improvement of the ability to predict male fertility increases the industry’s profit by improving the conception rate of herds.

Artificial insemination is the general method of breeding for genetic improvement in offspring. However, almost half of the insemination cases fail to achieve full term pregnancy; due to male infertility or subfertility. To maximize the success of insemination, accurate semen quality testing is required prior to insemination.

But the quantitative parameters such as sperm count, motility; and viability are the first step of analysis for identifying male factor infertility. Although these parameters are well standardizing as routine semen analysis methods worldwide; the clinical value is still questionable. But to improve the accuracy of male fertility prediction, our group developed and optimized the sperm penetration assay (SPA) in porcine and bull spermatozoa; which representing more than 95% accuracy for predicting male fertility.

Combination of multiple biomarkers

Even though basic semen analyses have been used to provide preliminary information, it cannot fully identify the superior or inferior fertility bulls. Therefore, more powerful and easy to use methods for the prediction of male fertility are required; such as proteomic or microarray chips. During past decades, omics approaches have been developing and suggesting the numerous fertility relating potential biomarkers.

The study identifying the fertility related protein markers, enolase1, ATP synthase, H+ transporting, mitochondrial F1 complex, beta subunit, voltage dependent anion channel 2, phospholipid hydroperoxide glutathione peroxide; and ubiquinol cytochrome c reductase complex core protein 2 in bovine spermatozoa. In the present study, we perform a marker combination assay using the western blot data of ENO1, ATP5B, VDAC2, GPx4, and UQCRC2 from 20 individual bull semen samples.

Fertility marker candidate

Although no meaningful changes existed in overall accuracy (70–85%) to discriminate the normal and below normal fertility between ENO1 single marker and combined marker panels; multiple marker combination methods using ENO1, VDAC2, GPx4, and UQCRC2 provided absolute sensitivity and NPV, with higher specificity (70%) and PPV (77%).

ENO1 can be used as a fertility marker candidate; but there were limitations for providing absolute information about normal and below normal fertility. Although the combined use of fertility related markers cannot provide absolute accuracy, it can help in indicating below-normal fertility in bulls. These results may contribute to the maintenance cost in the animal industry; via selection of bulls with inferior fertility.