Objective: To investigate the application of the self-made semen quality control (QC) product in internal QC of computer-assisted sperm analysis (CASA). METHODS: CASA was calibrated with high- and low-concentration commercially available semen QC product and meanwhile 15 samples of self-made mixed semen QC product were placed in 75 cryotubes containing liquid nitrogen, followed by CASA of the concentration, motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), wobble (WOB) and straightness (STR) of the sperm using standard procedures and 50 days of continuous monitoring. The Makler counting plate was used to measure the concentration and motility of the self-made sperm. RESULTS: The coefficients of variation (CV) of the commercially available semen QC product at high and low concentrations were 6.18% and 7.85%, respectively. CASA showed that the concentration of the self-made QC product was (25.97 ± 1.41) ×10⁶/ml, with a CV of 5.42%, and the sperm motility, VCL, VSL, VAP, LIN, WOB and STR were (22.15 ± 1.75)% (CV = 7.9%), (59.18 ± 2.05) μm/s (CV = 3.46%), (26.79 ± 1.2) μm/s (CV = 4.48%), (34.98 ± 1.4) μm/s (CV = 4.01%), 46.81 ± 1.55 (CV = 3.3%), 60.52 ± 1.3 (CV = 2.15%) and 76.46 ± 1.98 (CV = 2.59%), respectively. The concentration and motility of the self-made sperm detected with the Makler counting plate were (34.39 ± 2.37) ×10⁶/ml (CV = 6.89%) and (38.04 ± 1.69)% (CV = 4.44%), respectively. Levey-Jennings QC charts were plotted for the indicators using the means and standard deviation. CONCLUSIONS The self-made internal QC product by liquid nitrogen cryopreservation is feasible and effective for monitoring the accuracy and precision of CASA-derived sperm concentration and motion parameters, and it has a smaller CV than the commercially available QC product in measuring sperm concentration.
Computers ; Humans ; Male ; Quality Control ; Semen Analysis/standards* ; Sperm Motility ; Spermatozoa

According to the World Health Organization (WHO), oxidative stress (OS) is a significant contributor to male infertility. Seminal OS can be measured by a number of assays, all of which are either costly or time sensitive and/or require large semen volume and complex instrumentation. One less expensive alternative is to quantify the oxidation-reduction potential (ORP) with the MiOXSYS. In this international multi-center study, we assessed whether ORP levels measured by the MiOXSYS could distinguish semen samples that fall within the 2010 WHO normal reference values from those that do not. Semen samples were collected from 2092 patients in 9 countries; ORP was normalized to sperm concentration (mV/10⁶ sperm/ml). Only those samples with a concentration >1 × 10⁶ sperm ml^-1 were included. The results showed that 199 samples fell within the WHO normal reference range while the remaining 1893 samples did not meet one or more of the criteria. ORP was negatively correlated with all semen parameters (P < 0.01) except volume. The area under the curve for ORP was 0.765. The ORP cut-off value (1.34 mV/10⁶ sperm/ml) was able to differentiate specimens with abnormal semen parameters with 98.1% sensitivity, 40.6% specificity, 94.7% positive predictive value (PPV) and 66.6% negative predictive value (NPV). When used as an adjunct to traditional semen analysis, ORP levels may help identify altered functional status of spermatozoa caused by OS in cases of idiopathic male infertility and in male partners of couples suffering recurrent pregnancy loss, and thereby directing these men to relevant medical therapies and lifestyle modifications.
Adult ; Area Under Curve ; Humans ; Infertility, Male/metabolism* ; Male ; Middle Aged ; Oxidation-Reduction ; Oxidative Stress ; ROC Curve ; Reference Values ; Semen/metabolism* ; Semen Analysis/standards* ; Sensitivity and Specificity ; Sperm Count/standards* ; Spermatozoa/metabolism* ; Young Adult

ObjectiveTo investigate the relationship of the characteristics of sperm donors with the results of screening and provide some reference for the screening of sperm donors.

METHODSWe statistically analyzed the screening data about 12 362 sperm donors at the Guangdong Human Sperm Bank from January 2003 to June 2017 and the relationship of the eligibility rate of screening with the donors' age, education, occupation, marriage, and fatherhood.

RESULTSOf the 12 362 sperm donors, 3 968 (32.1%) met the standards of semen quality and 3 127 (25.3%) filled all the requirements of sperm donation. The eligibility rate of screening was 27.7% in the donors aged 20－24 years, 24.3% in those aged 25－29 years, 23.8% in those aged 30－34 years, and 17.5% in those aged =≥35 years (P < 0.01); 23.5% in the senior high school students, 24% in the junior college students, 25.9% in the undergraduates, and 30.3% in the postgraduates (P < 0.01); 29.3% in the students versus 22.9% in the others (P < 0.01), 41.5% in the married versus 20.7% in the unmarried (P < 0.01), and 45.6% in the fathers versus 20.9% in the childless husbands (P < 0.01).

CONCLUSIONSA higher eligibility rate of screening was found among the sperm donors aged <35 years or with a bachelor's or higher degree, particularly among students.

Adult ; Humans ; Male ; Semen Analysis ; standards ; Spermatozoa ; Students ; statistics & numerical data ; Tissue Donors ; statistics & numerical data ; Tissue and Organ Procurement ; Young Adult

Objective: To investigate the association of the abnormal length of human Y chromosome with semen quality and the outcome of assisted reproductive technology (ART). METHODS: Based on the karyotype, we assigned the patients undergoing ART to a normal control, a long Y chromosome (Y>18), and a short Y chromosome group (Y<22). We compared the semen parameters and numbers of embryos and high-quality embryos among the three groups of patients and performed statistical analysis of the obtained data using Chi-square distribution and t-test. RESULTS: Compared with the control, the Y>18 group showed a significantly lower incidence rate of asthenozoospermia (31.03% vs 8.33%, P <0.05) and a larger number of high-quality embryos (5.46 ± 4.54 vs 7.40 ± 5.49, P<0.05). Both the incidence rate of azoospermia and number of total embryos were remarkably lower in the control than in the Y<22 group (1.87% vs 16.47%, P <0.05; 8.60 ± 7.03 vs 10.00 ± 6.58, P<0.05). No statistically significant differences were found in the pregnancy rate between the Y>18 and Y<22 groups (P>0.05). CONCLUSIONS Short Y chromosome may affect spermatogenesis, but the length of Y chromosome does not negatively influence the outcome of ART.
Asthenozoospermia ; genetics ; Azoospermia ; genetics ; Chi-Square Distribution ; Chromosomes, Human, Y ; Female ; Humans ; Karyotype ; Karyotyping ; Male ; Pregnancy ; Pregnancy Rate ; Reproductive Techniques, Assisted ; Semen ; Semen Analysis ; standards ; Sex Chromosome Aberrations ; Spermatogenesis ; Treatment Outcome

In recent years, the variation trend of male fertility and semen parameters has aroused much academic controversy and become a focus of public attention. For the assessment of male fertility, female pregnancy is regarded as a gold standard, but semen parameters are commonly used as surrogate or indirect evidence in clinical practice and laboratory research. The reference range of se- men parameters being used in China is based on the WHO recommended data and lacks the specific reference value for healthy Chinese men. No definite conclusion has yet been derived from studies at home and abroad on the general variation trend of semen parameters worldwide, but many researchers agree on the decline of semen quality in some areas of the world. Long-term continuous prospective studies are needed for the evaluation and prediction of the general variation trend of semen quality.
Asian Continental Ancestry Group ; China ; Female ; Fertility ; Humans ; Male ; Pregnancy ; Prospective Studies ; Reference Values ; Semen ; physiology ; Semen Analysis ; standards ; Sperm Count ; World Health Organization

OBJECTIVETo determine the predictive value of sperm morphology based on the criteria of the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO5) on the outcomes and neonatal status following IVF-ET.

METHODSAccording to the strict criteria of WHO5, we obtained semen samples from 789 subjects and allocated them to a normal group (morphologically normal sperm > or = 4%, 754 cycles) and a teratozoospermia group (morphologically normal sperm < 4%, 35 cycles). We made comparisons between the two groups in the rates of normal fertilization, cleavage, quality embryo, implantation, clinical pregnancy and miscarriage as well as the status of the neonates.

RESULTSNo significant differences were observed in the couples' age, mean number of oocytes, and mean stature and body mass index of the women between the two groups (P > 0.05). The teratozoospermia group showed slightly lower rates of fertilization, cleavage, quality embryo, embryo cryopreservation, implantation and pregnancy, but a higher rate of miscarriage than the normal group (P > 0.05). Apart from 141 on-going pregnancies (140 in the normal and 1 in the teratozoospermia group), 228 healthy infants were born following 789 transfer cycles, 213 (141 singletons and 36 twins) in the former and 15 (9 singletons and 3 twins) in the latter group. Congenital defects were found in none of the neonates, and there were no significant differences in the gestation period, premature birth rate and low body weight between the two groups (P > 0.05).

CONCLUSIONSperm morphology according to the criteria of WHO5 has but a limited value in predicting the outcomes and neonatal status following IVF-ET.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infant, Newborn ; Infertility, Female ; therapy ; Male ; Predictive Value of Tests ; Pregnancy ; Pregnancy Outcome ; Reference Standards ; Semen Analysis ; Spermatozoa ; Treatment Outcome

OBJECTIVETo establish a method for internal quality control (IQC) of sperm concentration test in the laboratory.

METHODSWe set the concentrations of frozen semen at 20 x 10(6) and 80 x 10(6) as low and high concentrations of putative IQC products, with QC-BEADSTM quality control beads (QCBs) as the control. Using the double-blind method, four technicians determined the sperm concentrations of the IQC products and QCBs by computer-assisted sperm analysis, and drew a quality control chart (Xbar chart and Sbar chart) for each product. Through a month of continuous detection, we calculated and compared the intra- and inter-batch coefficients of variation (CV%) of the quality control products of high and low concentrations.

RESULTSThe intra-batch coefficients of variation of the assumed IQC products of high and low concentrations were CV3.5% and CV2.4%, and their inter-batch coefficients of variation were CV10.2% and CV9.6%. The intra-batch coefficients of variation of the QCBs of high and low concentrations were CV5.1% and CV7.1%, and their inter-batch coefficients of variation were CV7.1% and CV8%. The intra-batch coefficients of variation of both IQC products and QCBs of high and low concentrations were <10%, and their inter-batch coefficients of variation were <15%, which conformed to Levey-Jennings quality control principles and achieved IQC purposes. No significant differences were found in either intra- or inter-batch coefficients of variation between the IQC products and QCBs of high and low concentrations (P>0.05), indicating that assumed IQC products can replace QCBs for internal quality control in the laboratory.

CONCLUSIONThe IQC method we established for determining sperm concentration is simple, feasible and reliable.

Double-Blind Method ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Semen Preservation ; Sperm Count ; Sperm Motility ; Spermatozoa

OBJECTIVETo establish the methods for training green-hand laboratorians in standard semen analysis and evaluating the training results, and afford some reference for internal quality control and training in andrology laboratories in China.

METHODSWe trained the green-hand technicians in standard semen analysis recommended by WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed), and evaluated the training results by assessing the trainees' performance in the examination of sperm concentration and motility by statistic analysis.

RESULTSAfter training, the coefficients of variation of the semen samples with high, middle and low sperm concentration achieved by the trainees were 7.72% and 3.38% and 4.49%, and those with high, middle and high motility were 7.82%, 8.09% and 6.62%, respectively. We used Bland-Altman's method to evaluate the consistency between the results obtained by the trainees and those by the trainers. For sperm concentration, 4.77% of the datum points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 8 x 10(6)/ml within the 95% consistency interval. For sperm motility, 7.15% of the data points were out of the 95% consistency interval, and the absolute value of the biggest difference between the trainees and trainers was 10% within the 95% consistency interval. Two-way analysis of variance showed no significant differences in the results of sperm concentration and motility analyses between the trainees and trainers (P > 0.05).

CONCLUSIONTraining in standard semen analysis significantly improved the precision of semen analysis among the green-hand laboratorians. The training and assessment methods we established proved to be effective and feasible.

Humans ; Inservice Training ; Laboratory Personnel ; education ; Male ; Quality Control ; Semen Analysis ; standards

OBJECTIVETo compare the criteria of sperm morphology evaluation in the fifth edition of WHO Laboratory Manual for the Examination and Processing of Human Semen and those in the fourth edition, and to know the changes in the criteria of sperm morphology evaluation in the new edition.

METHODSNine technicians from Zhejiang Human Sperm Bank evaluated the morphology of 1 000 spermatozoa in 96 sperm morphological pictures according to the criteria in the fourth and fifth editions of WHO Laboratory Manual, respectively.

RESULTSThe percentage of morphologically normal sperm by the criteria of the fifth edition was (26.50 +/- 5.06)%, significantly higher than (11.39 +/- 3.17)% by the fourth edition (P < 0.05), while the rates of sperm head and tail defects based on the former were (64.26 +/- 7.66)% and (10.92 +/- 2.03)%, significantly lower than (76.11 +/- 8.18)% and (39.89 +/- 3.85)% according to the latter (P < 0.05). There were no significant differences in the rates of sperm midpiece defects and excessive residual cytoplasm by the fifth and fourth editions ([16.46 +/- 3.08]% vs [15.22 +/- 3.51 ]% and [4.24 +/- 1.66]% vs [3.87 +/- 1.68]%, P > 0.05).

CONCLUSIONThe criteria of sperm morphology evaluation in the fifth edition of WHO Laboratory Manual are less strict than those in the fourth, and the percentage of morphologically normal sperm is higher according to the fifth edition.

Humans ; Male ; Semen Analysis ; standards ; Sperm Head ; ultrastructure ; Sperm Midpiece ; ultrastructure ; Sperm Motility ; Spermatozoa ; ultrastructure ; World Health Organization

OBJECTIVETo investigate the effects of the computer-assisted semen analysis (CASA) on human sperm movement parameters at different times after semen collection.

METHODSNinety-two semen samples with sperm density > or = 20 x 10(6)/ml and sperm liquefaction time < 20 min were placed in a incubation box at the temperature of 37 degrees C. Then the seminal parameters were analyzed with the computer-assisted semen analysis (CASA) system at 20, 30, 60 and 90 min after semen collection.

RESULTSThe percentages of grade a and b sperm were significantly lower at 30, 60 and 90 min than at 20 min (P < 0.05), so were the percentages of grade c sperm at 60 and 90 min than at 20 and 30 min (P < 0.05), but there were no significant differences in the percentage of grade c sperm between the 20-min and 30-min groups (P > 0.05). The percentages of grade a + b and a + b + c sperm were also significantly lower at 30, 60 and 90 min than at 20 min (P < 0.05). The beat cross frequency (BCF) was significantly higher at 30 min than at 20 min (P < 0.05), while the lateral head amplitude (ALH) significantly lower at 90 min than at 30 min (P < 0.05). The sperm wobbliness (WOB) was significantly higher while the curvilinear velocity (VCL) significantly lower at 90 min than at 20 and 30 min (P < 0.05). Straightness (STR) at 30, 60 and 90 min, and average path velocity (VAP) and straight line velocity (VSL) at 90 min were significantly lower than at 20 min (P < 0.05). There were no significant differences in sperm density, average motion degree (MAD) and linearity (LIN) among the four groups (P > 0.05).

CONCLUSIONThe interval between semen collection and sperm routine analysis needs to be standardized. The results of this study suggest that sperm movement parameters of normal liquefied semen samples are relatively constant at 30 -60 min after semen collection.

Adult ; Humans ; Male ; Reference Standards ; Semen Analysis ; Sperm Count ; Sperm Motility ; Time Factors