The semen analysis is one, if not the most, important and widely used clinical laboratory test to evaluate the fertility potential of the male. However, recent reports have suggested that the semen analysis is unreliable. Quality control in the andrology laboratory is often seen as problematic, and many laboratories do not routinely employ QC procedures in semen testing. Quality assurance is an often overlooked and unappreciated aspect of overall quality laboratory performance. External proficiency testing programs in andrology are not universally accepted, and the results from the few programs currently available demonstrate huge variations between laboratories. Numerous different standards and criteria are being used by andrology laboratories, making it difficult if not impossible to compare results from one laboratory to another. However, reliable semen analyses can be obtained by following several recommendations: (1) all laboratories performing the semen analysis should adopt universally accepted performance standards and criteria, (2) all laboratories performing this test should participate in external proficiency testing programs, (3) andrology laboratories should implement effective internal quality control and quality assurance programs to ensure that the results reported are accurate and reproducible, and (4) physicians should only refer their patients to, or accept semen analysis results from, laboratories that have stringently followed these recommendations.
Clinical Laboratory Techniques ; standards ; Humans ; Male ; Quality Control ; Reproducibility of Results ; Semen ; Sperm Count ; standards ; Sperm Motility

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

OBJECTIVETo discuss how some pre-analysis processes influence the results of semen analysis and how to minimize their influence on the accuracy of laboratory results based on the concept of total quality management (TQM).

METHODSWe conducted semen quality analyses for 21 male volunteers, who had abstained from tobacco and alcohol for over 72 days for the purpose of fertilization, before and after the abstinence, and obtained their seminal parameters at 0.5, 1, 2 and 3 hours after semen sample collection.

RESULTSSperm concentration, sperm motility and the percentage of grade a + b sperm were significantly higher after the abstinence of tobacco and alcohol than before (P < 0.01). With the lengthening of post-ejaculation time, there was a significant decrease in sperm motility and the percentage of grade a + b sperm (P < 0.05), but not in sperm concentration (P > 0.05).

CONCLUSIONA lot of factors may affect the results of semen analysis, including the subjects' habits of drinking and smoking and the length of time after semen collection. Therefore, every procedure of semen analysis has to be dealt with very carefully so as to meet the requirements of TQM and achieve most reliable results for clinical use.

Adult ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Smoking Cessation ; Sperm Count ; Sperm Motility ; Temperance

Semen analysis is a basic test to evaluate male reproductive function. In recent years, urgent needs for the standardization of semen analysis have been emphasized among andrologists worldwide. This review discusses the standardization and quality control (QC) for the analysis of sperm quality parameters, including sperm concentration, motility and morphology. The key to sperm concentration analysis is the standardization of sperm-counting chamber, thus Cell-VU chamber may be the first choice. The analysis of sperm motility and morphology is too subjective to be reliable. Therefore, the computer-aided semen analysis (CASA) system may be the final selection. QC of semen analysis mainly lies in the selection of QC materials and the administration of external QC and internal QC. Meanwhile, the charts and arithmetic methods should be established to monitor QC of semen analysis.
Humans ; Male ; Quality Control ; Reference Standards ; Semen ; cytology ; Sperm Count ; instrumentation ; standards ; statistics & numerical data ; Sperm Motility ; Spermatozoa ; cytology ; physiology

Clinical Laboratory Techniques ; standards ; Humans ; Male ; Quality Control ; Semen ; Sperm Count ; methods ; standards

OBJECTIVESemen evaluation is the most important laboratory test for assessing male fertility. However, lack of strict quality control (QC) for semen analyses in hospital andrology laboratories makes it difficult and meaningless to compare semen data between different laboratories. This paper reports a comparative study on the accuracy of the Hemacytometer (Qiujing Inc., Shanghai, China), Makler (Sefi-Medical Instrument, Haifa, Israel), and Cell-VU (Millennium Sciences Inc., New York, USA) chambers for sperm counting.

METHODSBoth low [(18 +/- 2.5) x 10(6)/ml] and high [(35 +/- 5) x 10(6)/ml] pre-calibrated standard latex bead solutions (Hamilton Thorne Biosciences, USA) were used as the quality control solution to perform counts on the three different counting chambers. Bead counts for the three different chambers were compared, and variability within the chambers determined for standard solutions at low and high concentrations of latex beads, respectively.

RESULTSMean bead concentrations for the Cell-VU, Hemacytometer and Makler chambers were (37.63 +/- 4.89), (42.74 +/- 4.98) and (53.52 +/- 6.67) x 10(6)/ml respectively for a standard solution containing (35 +/- 5) x 10(6) beads/ml, and (18.22 +/- 1.77), (20.48 +/- 1.56), (24.97 +/- 4.75) x 10(6)/ml respectively for a standard solution containing (18 +/- 2.5) x 10(6) beads/ml. Mean bead concentrations for the Cell-VU chamber were consistently similar and close to the standard pre-calibrated bead solutions, while those for both the Hemacytometer and the Makler chambers were significantly overestimated (P < 0.001). The average coefficients of variation for the Cell-VU chamber were 7.51% for a higher concentration of the standard solution containing (36 +/- 5) x 10(6) beads/ml and 1.22% for a lower concentration of the standard solution containing (18 +/- 2.5) x 10(6) beads/ml, while the mean variation rates of the Hemacytometer and Makler chambers were 22.11% and 13.78% for a standard solution containing (36 +/- 5) x 10(6) beads/ml, and 52.91% and 38.72% for a standard solution containing (18 +/- 2.5) x 10(6) beads/ ml, respectively.

CONCLUSIONSemen analysis is one of the most important tests for male fertility evaluation, but the data obtained from commercially available counting chambers may differ markedly in accuracy and reliability. Results from this comparative study demonstrated that the Cell-VU chamber exhibits significantly more accurate and less variable results than those of the Hemacytometer and Makler chambers. To ensure the best possible evaluations and accurate diagnoses, we therefore recommend that Cell-VU be used as the standard counting chamber for routine semen analyses in andrology laboratories.

Blood Cell Count ; instrumentation ; Humans ; Male ; Quality Control ; Sperm Count ; instrumentation ; standards

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 investigate and analyze the results of the determination of sperm concentration, fructose concentration, alpha-glucosidase and acid phosphatase (ACP) activities in the seminal plasma from different hospitals in the city of Nanjing, so as to provide a basis for the external quality control (EQC) of semen analysis within Jiangsu Province or even the whole country.

METHODSEight samples of quality control products for low and high concentrations sperm count, fructose, alpha-glucosidase and ACP determination were prepared and divided, each detected for the sperm concentration, fructose, alpha-glucosidase and ACP activity, and the coefficient variances (CVs) were calculated. The products were then distributed to 11 hospitals in the city, and the results were collected and analyzed. In addition, the total relative errors (REs) for each product was calculated based on the results after dividing as reference values.

RESULTSThe CVs from the 8 samples after dividing were 3.83% - 11.16%. Collected from the 11 hospitals attending EQC were 11 reports of the results of sperm concentration, and 5 the results of fructose, alpha-glucosidase and ACP in seminal plasma. Among the results from different laboratories, those of fructose determination showed the minimal difference (CVs: 8.99% and 3.95% for low and high concentrations, respectively) , next came alpha-glucosidase (CVs: 16.66% and 18.41% for low and high activities, respectively), and ACP determination showed the maximal difference (CVs: 54.12% and 65.58% for low and high activities, respectively). Moreover, the same trend was observed in RE values, as shown in the total REs, which were 11.99% (low concentration) and 20.31% (high concentration) for the determination of fructose in seminal plasma, 22.92% and 27.26% for alpha-glucosidase, 7.34% and 318.35% for ACP in different laboratories, and the maximal RE value was detected in the result of the high-activity ACP sample. Of the 11 hospitals, 6 determined sperm concentration with the computer-assisted semen analysis (CASA) system, and the other 5 with the modified hemocytometer. RE values (148.47% and 187.59% for low and high concentration samples, respectively) and sperm concentrations ([62.74 +/- 16.63] x 10(6)/ml and [163.32 +/- 36.24] x 10(6)/ml) counted with the hemocytometer were significantly higher than those with the CASA system (REs 13.97% and 10.48%; sperm concentrations [24.88 +/- 4.16] x 10(6)/ml and [54.24 +/-23.06] x 10(6)/ml ).

CONCLUSIONThe methods of seminal alpha-glucosidase and fructose determination were relatively stable in current andrology laboratories, and the variance range could be accepted. However, the method of seminal ACP determination might be unadaptable to clinical application, and needs to be further improved. Hemocytometer, which significantly overestimated sperm concentration, could not be applied to the assay of sperm concentration.

Acid Phosphatase ; analysis ; China ; Humans ; Male ; Quality Control ; Semen ; enzymology ; Sperm Count ; standards ; Sperm Motility ; alpha-Glucosidases ; analysis

OBJECTIVETo investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures.

METHODSUsing alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method.

RESULTSThree agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05).

CONCLUSIONThe optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

Asthenozoospermia ; genetics ; Comet Assay ; standards ; DNA Damage ; DNA Fragmentation ; Humans ; Hydrogen Peroxide ; toxicity ; Male ; Oligospermia ; genetics ; Oxidants ; toxicity ; Reproducibility of Results ; Sperm Count ; Spermatozoa ; drug effects ; enzymology ; Time Factors

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