Correlation Analysis of the Results of Double Fluorescence (AO/PI) Staining and Clinical Outcomes

ObjectiveTo estimate the predictive value of double fluorescence [acridine orange (AO)/propidium iodide (PI)] staining results for fertilization rate and clinical outcomes and analyze the correlation between the results of AO/PI staining and sperm apoptosis.MethodsA prospective study was carried out, 235 infertile couples remedied using traditional in vitro fertilization (IVF) were included. Semen collected from 235 patients were stained by fluorescence dye, AO and PI, at the same time the spermatozoa apoptosis rate was calculated by using flow cytometry to detect the rate of Annexin V+/PI−. The result of fluorescence was divided into 3 groups as green (G), yellow (Y) and red (R) according to the color of fluorescence. The correlation between the percent of the 3 colors and clinical outcomes and spermatozoa apoptosis rate was evaluated.ResultsSignificant negative correlation was observed between the percentage of Y and fertilization rate (r=-0.42, P=0.04), no significant correlation was observed between the percentage of G, R and fertilization rate. The percentage of G, Y, R was not significantly different between pregnant patients and non-pregnant patients, respectively, while the percentage of Y was significant different between miscarriage patients and liveborn patients (r=0.61, P=0.01) and no significant difference exist in the percentage of G and R between liveborn patients and miscarriage patients. There was a significant positive correlation between the percentage of Y and the rate of Annexin V+/ PI− (r=0.53, P=0.04), and no significant correlation was shown between the percentage of G, R and the rate of Annexin V+/ PI−.ConclusionsThe percentage of yellow group of double fluorescence (AO/PI) staining will affect the fertilization rate and the miscarriage rate, and this group of spermatozoa may be connected with the spermatozoa apoptosis.