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SCSA Diagnostics is the pioneer in sperm DNA analysis, leveraging established research and ongoing clinical studies for over 25 years to perform one of the most advanced sperm DNA fragmentation tests available.
The development of the SCSA test was presented in the renowned journal Science in December 1980.
Note that there are some labs that offer a SCSA-like test. However, it is not the same precision SCSA ® test.
Fragmentation of sperm DNA is one of the most important paternal causes of repeated reproductive failure. About 20-30% of men who attend a fertility clinic have a level of sperm DNA fragmentation that can lead to pregnancy failure.
Spano et al. 2000 The dashed line shows the clinical threshold for sperm DNA fragmentation and pregnancy chances
Some authors have published that semen samples containing more than 27-30% sperm with DNA fragmentations (DNA Fragmentation Index, %DFI) are 7-10 times less likely to get pregnant through intercourse or IUI, and one around 1.5 -2.0 times lower chance through IVF/ICSI
Giwercman A et al. (2009 in press, Int. J. Androl) included 137 men from couples with proven infertility with no known maternal factors and 137 men with proven fertility
In men with normal semen parameters (sperm concentration, motility and morphology), the risk of infertility was increased 5-fold with DFI values above 20%.
However, if any of the semen parameters were abnormal, the risk of infertility was increased 16-fold even with DFI values above 10%
The authors concluded that “DFI is a predictor of the likelihood of natural conception, even when classic semen parameters are within the normal range. The SCSA analysis complements treatment planning for couples with proven infertility.”
The integrity of sperm DNA is continually challenged by endogenous and exogenous factors, although various mechanisms for repairing and protecting DNA from damage are active in human cells (Hoeijmakers, 2009). In particular, the preservation of DNA integrity plays a major role in passing the genome on to the next generation. In these cells, DNA double-strand breaks are physiologically induced during spermatogenesis and spermiogenesis to facilitate meiotic crossover and histone-protamine substitution, respectively (Rathke et al., 2014). Apart from this “physiological” DNA damage, other exogenous and endogenous factors can affect DNA integrity during sperm maturation/storage in the epididymis (Moustafa et al., 2004; Ramos et al., 2004; Sakkas et al., 2002) . Studying DNA integrity could be a fundamental step in assessing sperm functional competence (Lewis et al., 2008). Therefore, analysis of sperm DNA damage could be crucial for both diagnosing infertility and predicting ART success. In the context of IVF, this assessment is of particular importance as it bypasses the natural selection barriers of conception, increasing the likelihood that sperm with significant DNA damage will transmit the genetic aberrations to the newborn (Host et al., 2000). Therefore, several studies have evaluated the predictive role of sperm DNA damage for either ART outcome or sperm selection. Sperm DNA damage can result from five different pathogenic mechanisms (Perrin et al., 2011; Sakkas and Alvarez, 2010).
DFI (DNA Fragmentation Index) :
DNA Fragmentation Index (%DFI) is the measure of sperm DNA fragmentation and an indicator of sperm quality
HDS (High DNA Stainability Cells): Percentage of sperm with abnormal core proteins and chromatin structures (immature spermatozoa)
The SCSA® test is performed using a laser-based device called a flow-cytometer which measures 250 sperm/second. The test result is based on the measurment of around 10,000 sperm.
SCREENING TEST : Particularly suitable for couples with a long-awaited desire to have children
UNSUCCESSFUL IUI/IVF/ICSI ATTEMPTS
REPEATED MISSORS
POOR EMBRYO DEVELOPMENT AND QUALITY
UNEXPLAINED BARRENESS: Couples who want to have children and have unremarkable diagnostic tests
MEN PRONE TO HIGH OXIDATIVE STRESS:
LIFESTYLE: Smoking, drug use, increased alcohol consumption, obesity, stress
IMPACT NUTRITION: A detailed analysis and appropriate modification of the diet can bring about a drastic improvement in sperm DNA fragmentation
HEAT STRESS: e.g. B. by whirlpools, working with a laptop on the lap, fever etc.
OCCUPATIONAL EXPOSURES: Chemicals, pesticides, fertilizers, dyes, etc.
DIABETES: An independent risk factor for increased sperm DNA fragmentation
MEDICATIONS: Even safe medications can cause sperm DNA fragmentation
ENVIRONMENTAL TOXINS / Air pollution
VARICOCELE: Once the diagnosis has been made, either by exam or ultrasound, the SCSA test can help in deciding whether or not to have surgery.
CANCER AND CANCER TREATMENTS chemotherapy and radiotherapy
CHRONIC DISEASES: Often cause increased oxidative stress leading to DNA damage
ANDROLOGICAL DISEASES / Genitourinary Tract Infections and Leukocytospermia: Local infection/inflammation causes direct damage to sperm DNA.
AGE >40 YEARS: Studies show that the older the man, the higher the DNA fragmentation
LUBRICANT: causes direct damage to sperm DNA
The SCSA® test is performed using a laser-based device called a flow cytometer. Essentially, the flow cytometer measures 250 sperm/s with high precision and provides a statistically robust result based on data from 2x 5000 sperm per sample. The precision of flow cytometry coupled with the precise biochemistry of the SCSA test enables highly accurate measurements of sperm chromatin and DNA integrity.
The cells are stained with a fluorescent DNA dye (acridine orange) and forced through a glass channel in a liquid suspension. When the cells pass the laser beam, the laser light causes the dye to emit fluorescent light of a specific color.
In the SCSA® test, due to the unique nature of the Acridine Orange dye, sperm that emit green fluorescence have undetectable levels of fragmented DNA, while sperm that emit yellow to red florescence have moderate to high levels of fragmented DNA.
Male fertility is traditionally examined using sperm tests (spermograms) that record sperm concentration, motility and morphology. However, numerous studies over the last half century clearly show that these parameters do not predict pregnancy, since fertile and infertile men have overlapping values (with the exception of azoospermia). The classic spermiograms are light microscopic measurements of external sperm factors, while the SCSA test measures the internal core factors that cannot be measured using the light microscope.
The SCSA test is a highly accurate test that requires a flow cytometer that is inaccessible to most fertility clinics due to high capital costs and a trained staff. Therefore, semen samples collected from you or a patient’s home can be cryopreserved natively.
You can inform us by e-mail about a desired transport. We will then send you a pre-cooled dryshipper; the semen samples are stored and sent back to our laboratory.
The SCSA report will be available to you via a secure WEB site approximately 4 business days after the sample arrives at our laboratory.
Collection and processing of the samples:
It should be obtained after 24-48 hours of abstinence by masturbating in sterile plastic sample containers. After liquefaction (liquefaction) of the sample, the sperm concentration, motility and morphology are determined. Write down the parameters on the consignment note.
The sperm concentration must not fall below 500,000/ml . Otherwise, inconclusive results may occur.
Cryopreservation should take place no more than 1 hour after delivery , since the DFI can be falsified after a longer waiting period.
Cryopreservation of the samples:
Samples should be frozen as soon as possible after liquefaction to minimize exposure to air, which can cause damage caused by reactive oxygen species.
Prepare 2 cryovials (1-2mL vials): Label with patient’s name and date of sample collection.
Flash freezing :
Pipette 0.25mL or more of the liquefied native ejaculate into each cryovial.
Cryoprotectants is not required!
Place the tightly closed vials in an upright position in the supplied metal holder and place this directly in the pre-cooled cryogenic container. The container must be closed quickly.
The Dryshipper is closed with the supplied cable tie and placed in the plastic container, to which the dispatch note is attached so that it is clearly visible.
Note: If the sperm concentration is sufficient, less than 0.5 ml (two 0.25 ml aliquots) of ejaculate may be sufficient for the SCSA® test. To perform the test we need two 0.20mL aliquots with a concentration >0.5M/mL.
Please contact our laboratory if you have any questions about the required volume of a sample of a specific concentration.
Sending the samples:
We ship a pre-chilled dryshipper. Dryshippers store the liquid nitrogen in a kind of “sponge” inside the container and thus ensure the safe transport of your cryopreserved samples in the nitrogen gas phase. The low temperature in the container can be maintained for 7 days. It is therefore important to send it back as soon as possible using the shipping label provided.
In the space between the dryshipper and the shipping box is an envelope with a cable tie and an ampoule holder. It also contains a shipping label and a corresponding sleeve that must be attached to the container for return shipping.
The inside of the dryshipper is very cold (-196°C). To avoid frostbite, you must wear gloves when touching anything that is in the dryshipper.
The SCSA test divides spermatozoa into five populations:
Percentage of sperm with immature chromatin. HDS sperm exhibit less chromatin condensation, resulting in increased DNA stainability. Some studies have shown that if the %HDS is over 36%, no pregnancy has occurred (Menezo).
The DPI values are divided into four statistical categories and can provide the following forecasts:
< 15% DFI = very good to good pregnancy results
> 15% to < 25% DFI = Good to moderate pregnancy outcomes
> 25% to < 40% DFI = Moderate to poor pregnancy outcomes
40% DFI = Very poor pregnancy outcomes
All data are to be understood without maternal influencing factors.
Furthermore, the finding shows a graphical representation of the %DFI (DNA Fragmentation Index) and the %HDS (High DNA Stainability)
The graph shows that 76% of the measured sperm show no DNA fragmentation and 24% show moderate-high DNA fragmentation (DFI = 24%). It is possible to conceive naturally or with the help of an IUI.
Current data shows that the probability of a successful pregnancy is significantly lower when the proportion of sperm with fragmented DNA is > 25%. However, a DFI > 25% does not rule out pregnancy. The 25% threshold is a statistical threshold. If a man has a constant DFI of >25%, he is placed in a statistical group that has been shown in clinical trials to take longer to achieve natural pregnancy and more IVF cycles. An increased rate of spontaneous abortions is associated with this or pregnancy does not occur at all.
Assessing sperm DNA quality
Since DFI and HDS are within the normal range, pregnancy is possible naturally or by insemination (ignoring partner factors affecting fertility)
That means the DNA of the sperm shows a high stability
The DFI is greatly increased. This means that the sperm DNA is very fragile and tends towards DNA fragmentation
Stabilization of DNA can be achieved by:
It may be necessary to switch from IUI to IVF/ICSI to increase the chances of a positive pregnancy outcome
The histogram shows the DFI (DNA Fragmentation Index) measured in a semen sample.
DFI scores are classified into four statistical categories and can predict the following (ignoring partner factors affecting fertility):
< 15% DFI = Very good to good pregnancy outcomes
> 15% to < 25% DFI = Good to moderate pregnancy outcomes
> > 25% to < 40% DFI = Moderate to poor pregnancy outcomes
> 40% DFI = Very bad outcome of pregnancy
If the %DFI is above 25%, the current literature recommends:
The ovum is believed to be able to repair some level of DNA damage.
Moderate to high levels of DNA fragmentation (DFI) found in the red fluorescent sperm likely exceed the egg’s DNA repair capacity.
Marcos Mesequer et al. found that after egg retrieval, every 10% increase in DFI increased the odds of not getting pregnant by a factor of 1.31. In comparison, this effect was not observed when using donor oocytes, which indicates the higher repair capacity with regard to DNA fragmentation of young oocytes.
If the %DFI is high, we recommend:
If there is a risk factor, repeat SCSA testing is recommended 3 months after risk factor removal/correction to assess impact.
Examples:
Nicotine abstinence (84 days later):
Varicocele surgery (3 months later):
Cessation of pesticide exposure (3 months later):
The cost of the test is €295. It is currently not covered by insurance.
Sending your samples to our laboratory is free of charge.
For DFI values >30%, we offer you a second free control test, which should normally take place 3 months after the first test.
Fragmentation of sperm DNA is one of the most important paternal causes of repeated reproductive failure. About 20-30% of men who attend a fertility clinic have a level of sperm DNA fragmentation that can lead to pregnancy failure.
Spano et al. 2000 The dashed line shows the clinical threshold for sperm DNA fragmentation and pregnancy chances
Some authors have published that semen samples containing more than 27-30% sperm with DNA fragmentations (DNA Fragmentation Index, %DFI) are 7-10 times less likely to get pregnant through intercourse or IUI, and one around 1.5 -2.0 times lower chance through IVF/ICSI
Giwercman A et al. (2009 in press, Int. J. Androl) included 137 men from couples with proven infertility with no known maternal factors and 137 men with proven fertility
In men with normal semen parameters (sperm concentration, motility and morphology), the risk of infertility was increased 5-fold with DFI values above 20%.
However, if any of the semen parameters were abnormal, the risk of infertility was increased 16-fold even with DFI values above 10%
The authors concluded that “DFI is a predictor of the likelihood of natural conception, even when classic semen parameters are within the normal range. The SCSA analysis complements treatment planning for couples with proven infertility.”
Sperm DNA integrity is continuously challenged by endogenous and exogenous factors, although different mechanisms of repairing and protecting against this damage are active in human cells (Hoeijmakers, 2009). This is particularly relevant in germ cells, which have to preserve DNA integrity to pass the genome to the next generation. In these cells, DNA double-strand breaks are physiologically induced during spermatogenesis and spermiogenesis to facilitate meiotic crossover and histone– protamine substitution, respectively (Rathke et al., 2014). Apart from this first ‘physiological’ DNA damage, other exogenous and endogenous factors could affect DNA integrity during sperm maturation and storage in the epididymis (Moustafa et al., 2004; Ramos et al., 2004; Sakkas et al., 2002). Thus, DNA integrity is constantly at risk and its assessment could be a fundamental step in the evaluation of sperm functional competence (Lewis et al., 2008). Hence, sperm DNA damage evaluation could be crucial for both infertility diagnosis and prediction of ART success. In the setting of IVF this evaluation plays a peculiar role because the natural selection barriers of conception are bypassed, increasing the possibility of spermatozoa with significant DNA damage transmitting the genetic aberrations to the newborn (Host et al., 2000). Thus, several trials have evaluated the predictive role of sperm DNA damage for either ART outcome or sperm selection.
DFI (DNA Fragmentation Index) :
Percentage of sperm with DNA fragmentation
HDS (High DNA Stainability Cells): Percentage of sperm with abnormal core proteins and chromatin structures (immature sperm)
The SCSA® test is performed using a laser-based device called a flow-cytometer which measures 250 sperm/second. The test result is based on the measurment of around 10,000 sperm.
LIFESTYLE: smoking, drug use, increased alcohol consumption, obesity, stress
NUTRITION: Although commonly overlooked, we have often seen drastic improvement in sperm DNA fragmentation after a detailed analysis and correction of daily nutrition.
HEAT EXPOSURE: e.g. from whirlpool baths, working with a laptop on the lap, fever, etc.
OCCUPATIONAL EXPOSURES: chemicals, pesticides, fertilizers, dyes, etc.
DIABETES: An independent risk factor for increased sperm DNA fragmentation
MEDICATION: Even the most unassuming medications have sometimes seen to cause sperm DNA fragmentation
ENVIRONMENTAL TOXINS /air pollution: Although it may appear general, a keen look into exposure should be ruled out.
VARICOCELE: Once diagnosed either by examination or ultrasound, SCSA test can help in the decision if surgery will help.
CANCER AND CANCER TREATMENTS (chemotherapy and radiotherapy) : causes direct damage to the DNA of cells.
CHRONIC DISEASES: often cause increased oxidative stress leading to increased DNA damage
ANDROLOGICAL DISEASES / Infections of the urogenital tract and leukocytospermia: Local infection / inflammation causes direct damage to the DNA in sperm through release of ROS (Reactive Oxygen Species).
AGE >40 YEARS: It has been extensively studied that the older the man, the higher the DNA fragmentation
LUBRICANTS: cause direct damage to the sperm DNA
Sperm DNA damage can result from five different pathogenic mechanisms (Perrin et al., 2011; Sakkas and Alvarez, 2010).
The SCSA® test is performed using a laser-based device called a flow cytometer. Essentially, the flow cytometer measures 250 sperm/s with high precision, providing a statistically robust result based on data from 2x 5000 sperm per sample. The precision of flow cytometry coupled with the precise biochemistry of the SCSA test provides highly accurate measurements of sperm chromatin and DNA integrity.
The cells are stained with a fluorescent DNA stain (acridine orange) and then forced through a glass channel in a liquid suspension. When the cells pass the laser beam, the laser light causes the dye to emit fluorescent light of a specific color.
In the SCSA® test, due to the unique nature of the Acridine Orange dye, sperm that emit green fluorescence have undetectable levels of fragmented DNA, while sperm that emit yellow to red florescence have moderate to high levels of fragmented DNA.
The SCSA test is a two-fold simultaneous flow cytometry measure of
in thousands of sperm in a fresh or frozen/thawed semen sample.
The extent of DNA strand breaks and abnormality of chromatin structure are related to male factor fertility, including time to couple pregnancy, IVF embryo quality, miscarriage, or infertility.
Male fertility is classically addressed by semen tests that include sperm density, motility and morphology. However, numerous studies over the past half century clearly show that, except for absence of sperm, these parameters do not predict pregnancy since fertile and infertile men have overlapping values. The classical semen tests are light microscope measures of external sperm factors while the SCSA test are measures of the internal nuclear factors not visible by light
microscopy.
The SCSA test is a high precision test that requires a flow cytometer which due to the need of a high capital expense and a trained technician is not amenable to most infertility clinics. Thus, semen samples collected, as per detailed instructions, in a clinic or in a patients’ home, can be flash frozen in a LN 2 dry shipper tank.(See support protocol 1).
Clinics can then send us an email informing us. A pre-cooled LN 2 shipper is then sent to the clinic and the semen samples are flash frozen and the LN 2 shipper then returned to a SCSA diagnostic testing lab.
Some clinics send frozen semen samples in well insulated dry ice containers.
SCSA clinical data are then sent via a secure WEB site to the physician or patient that ordered the test.
There are five main populations of sperm identified by the SCSA® test:
the percentage of sperm with immature chromatin. HDS sperm exhibit less chromatin condensation, resulting in increased DNA stainability. Some studies have shown that if the %HDS is above 36%, no pregnancies have occurred (Menezo)
Our SCSA clinical report lists four statistical categories of fertility potential. These were derived from a comprehensive study of male fertility potential without medical intervention and have been confirmed in additional studies.
< 15% DFI = Excellent to good pregnancy outcomes (excluding female infertility factors)
> 15% to < 25% DFI = good to fair pregnancy outcomes
> 25% to < 40% DFI = fair to poor pregnancy outcomes
> 40% DFI = Very poor pregnancy outcomes
Furthermore, the finding shows a graphical representation of the %DFI (DNA Fragmentation Index) and the %HDS (high DNA stainability)
The graph shows that 76% of the measured sperm shows no DNA fragmentation and 24% shows medium-high DNA fragmentation (DFI= 24%). Pregnancy naturally or with the help of an IUI is possible.
Current data shows that the probability of a successful pregnancy is significantly lower when the proportion of sperm with fragmented DNA is > 25%. Nevertheless, a DFI > 25% does not rule out a normal pregnancy. The 25% threshold is a statistical threshold. Therefore, if a man has a constant DFI >25%, he will be placed in a statistical group of men who have been shown in clinical trials to take longer to achieve natural pregnancy and more IVF cycles. An increased rate of spontaneous abortions is associated with this or pregnancy does not occur at all.
The ovum is believed to be able to repair a certain amount of DNA damage.
Moderate to high levels of DNA fragmentation (DFI) found in the red fluorescent sperm likely exceed the egg’s DNA repair capacity.
Marcos Meseguer et al found that when oocytes from patients were taken, for every 10% increase in DFI, there was a 1.31 times increase in probability of not achieving pregnancy. Whereas, the same was not seen when donor oocytes were employed suggesting the higher capability of young oocytes to repair sperm DNA damage.
REPEATED IUI / IVF / ICSI FAILURE : The reason behind the unsuccessful attempts at IUI/IVF/ICSI can be identified and managed
REPEATED MISCARRAGE : An explanation for repeated spontaneous abortions (RPL, repeated miscarriage) could be found and guided
POOR EMBRYO DEVELOPMENT AND QUALITY : The reason behind poor embryo quality could be the fragmented sperm DNA which could be corrected before the next treatment
MEN AT RISK FOR HIGHER OXIDATIVE STRESS:INITIAL SCREENING TEST : Especially for couples who have been attempting pregnancy for a long time to identify the best management strategy
UNEXPLAINED INFERTILITY : Couples desiring children with all normal diagnostic tests
In cases of high DFI, we suggest:
When a risk factor is identified, after avoidance / correction of the risk factor a repeat SCSA test after 3 months is advised.
Yes. We offer a SCSA test at half the cost for controlling the effect of avoidance / correction of a risk factor.
Please find a few examples below:
After quitting smoking for 84 days
3 months after varicocele correction surgery :
3 months after stopping pesticide exposure:
After 24-48 hours of abstinence, the samples should be taken by masturbation into sterile plastic sample containers. After liquefaction of the sample, the sperm concentration, motility and morphology are determined.
Cryopreservation should take place no more than 1 hour after delivery, since the DFI can be falsely raised after a longer waiting period.
Flash-freezing: Freeze 0.25mL or more of liquefied native ejaculate in 1-2mL cryovials. Cryoprotectants is not required! (Please freeze 2 cryovials per patient)
Samples should be frozen as soon as possible after liquefaction to minimize exposure to air, which can cause damage caused by reactive oxygen species.Cryovials should be tightly capped, labeled with two identifiers (e.g. name and date of birth) and placed in an upright position directly into the pre-chilled cryo-container. The container must be closed quickly.
Note: Cryoprotectants are not needed since quick-frozen sperm give the clearest SCSA® data. This feature is unique to mammalian sperm cells due to the highly condensed, crystalline nature of the nuclei.
Note: If the sperm concentration is adequate, less than 0.5 mL (two 0.25 mL aliquots) of semen may be sufficient for the SCSA® test. To perform the test, we require two 0.20 mL aliquots with a concentration of >0.5 million/ml. Please contact our laboratory if you have questions pertaining to the volume necessary for a sample of a certain concentration.
We ship a pre-chilled dry shipper. Dry shippers store the liquid nitrogen in porous material and thus ensure the safe transport of your cryopreserved samples in the nitrogen gas phase.
The low temperature in the container can be maintained for 7 days. It is therefore important to send it back as soon as possible using the mailed/supplied shipping label.
Note: liquid nitrogen is adsorbed by special materials inside the shipping container. Even if they are turned upside down during transport, no liquid nitrogen will leak out. Dry shippers are not considered dangerous.
After receiving the sample(s) in our laboratory, we need 4 working days to create a report, which we will send to you.
The cost of the test is 295 Euro.
* Samples are shipped in a pre-cooled shipper that we can provide either at ambient or pre-chilled temperatures. The pre-cooled shipper will be sent to the centre where you have provided your sample for the centre to ship it to us.
The test at present is not covered by the insurance.
Please write us an email to contact@scsalab.com and after contacting you, we will send toy a dry shipper for the patient sample to be sent to us.
Fragmentation of sperm DNA is one of the most important paternal causes of repeated reproductive failure. About 20-30% of men who attend a fertility clinic have a level of sperm DNA fragmentation that can lead to pregnancy failure.
Spano et al. 2000 The dashed line shows the clinical threshold for sperm DNA fragmentation and pregnancy chances
Some authors have published that semen samples containing more than 27-30% sperm with DNA fragmentations (DNA Fragmentation Index, %DFI) are 7-10 times less likely to get pregnant through intercourse or IUI, and one around 1.5 -2.0 times lower chance through IVF/ICSI
Giwercman A et al. (2009 in press, Int. J. Androl) included 137 men from couples with proven infertility with no known maternal factors and 137 men with proven fertility
In men with normal semen parameters (sperm concentration, motility and morphology), the risk of infertility was increased 5-fold with DFI values above 20%.
However, if any of the semen parameters were abnormal, the risk of infertility was increased 16-fold even with DFI values above 10%
The authors concluded that “DFI is a predictor of the likelihood of natural conception, even when classic semen parameters are within the normal range. The SCSA analysis complements treatment planning for couples with proven infertility.”
Sperm DNA integrity is continuously challenged by endogenous and exogenous factors, although different mechanisms of repairing and protecting against this damage are active in human cells (Hoeijmakers, 2009). This is particularly relevant in germ cells, which have to preserve DNA integrity to pass the genome to the next generation. In these cells, DNA double-strand breaks are physiologically induced during spermatogenesis and spermiogenesis to facilitate meiotic crossover and histone– protamine substitution, respectively (Rathke et al., 2014). Apart from this first ‘physiological’ DNA damage, other exogenous and endogenous factors could affect DNA integrity during sperm maturation and storage in the epididymis (Moustafa et al., 2004; Ramos et al., 2004; Sakkas et al., 2002). Thus, DNA integrity is constantly at risk and its assessment could be a fundamental step in the evaluation of sperm functional competence (Lewis et al., 2008). Hence, sperm DNA damage evaluation could be crucial for both infertility diagnosis and prediction of ART success. In the setting of IVF this evaluation plays a peculiar role because the natural selection barriers of conception are bypassed, increasing the possibility of spermatozoa with significant DNA damage transmitting the genetic aberrations to the newborn (Host et al., 2000). Thus, several trials have evaluated the predictive role of sperm DNA damage for either ART outcome or sperm selection.
DFI (DNA Fragmentation Index) :
Percentage of sperm with DNA fragmentation
HDS (High DNA Stainability Cells): Percentage of sperm with abnormal core proteins and chromatin structures (immature sperm)
The SCSA® test is performed using a laser-based device called a flow-cytometer which measures 250 sperm/second. The test result is based on the measurment of around 10,000 sperm.
LIFESTYLE: smoking, drug use, increased alcohol consumption, obesity, stress
NUTRITION: Although commonly overlooked, we have often seen drastic improvement in sperm DNA fragmentation after a detailed analysis and correction of daily nutrition.
HEAT EXPOSURE: e.g. from whirlpool baths, working with a laptop on the lap, fever, etc.
OCCUPATIONAL EXPOSURES: chemicals, pesticides, fertilizers, dyes, etc.
DIABETES: An independent risk factor for increased sperm DNA fragmentation
MEDICATION: Even the most unassuming medications have sometimes seen to cause sperm DNA fragmentation
ENVIRONMENTAL TOXINS /air pollution: Although it may appear general, a keen look into exposure should be ruled out.
VARICOCELE: Once diagnosed either by examination or ultrasound, SCSA test can help in the decision if surgery will help.
CANCER AND CANCER TREATMENTS (chemotherapy and radiotherapy) : causes direct damage to the DNA of cells.
CHRONIC DISEASES: often cause increased oxidative stress leading to increased DNA damage
ANDROLOGICAL DISEASES / Infections of the urogenital tract and leukocytospermia: Local infection / inflammation causes direct damage to the DNA in sperm through release of ROS (Reactive Oxygen Species).
AGE >40 YEARS: It has been extensively studied that the older the man, the higher the DNA fragmentation
LUBRICANTS: cause direct damage to the sperm DNA
Sperm DNA damage can result from five different pathogenic mechanisms (Perrin et al., 2011; Sakkas and Alvarez, 2010).
The SCSA® test is performed using a laser-based device called a flow cytometer. Essentially, the flow cytometer measures 250 sperm/s with high precision, providing a statistically robust result based on data from 2x 5000 sperm per sample. The precision of flow cytometry coupled with the precise biochemistry of the SCSA test provides highly accurate measurements of sperm chromatin and DNA integrity.
The cells are stained with a fluorescent DNA stain (acridine orange) and then forced through a glass channel in a liquid suspension. When the cells pass the laser beam, the laser light causes the dye to emit fluorescent light of a specific color.
In the SCSA® test, due to the unique nature of the Acridine Orange dye, sperm that emit green fluorescence have undetectable levels of fragmented DNA, while sperm that emit yellow to red florescence have moderate to high levels of fragmented DNA.
The SCSA test is a two-fold simultaneous flow cytometry measure of
in thousands of sperm in a fresh or frozen/thawed semen sample.
The extent of DNA strand breaks and abnormality of chromatin structure are related to male factor fertility, including time to couple pregnancy, IVF embryo quality, miscarriage, or infertility.
Male fertility is classically addressed by semen tests that include sperm density, motility and morphology. However, numerous studies over the past half century clearly show that, except for absence of sperm, these parameters do not predict pregnancy since fertile and infertile men have overlapping values. The classical semen tests are light microscope measures of external sperm factors while the SCSA test are measures of the internal nuclear factors not visible by light
microscopy.
The SCSA test is a high precision test that requires a flow cytometer which due to the need of a high capital expense and a trained technician is not amenable to most infertility clinics. Thus, semen samples collected, as per detailed instructions, in a clinic or in a patients’ home, can be flash frozen in a LN 2 dry shipper tank.(See support protocol 1).
Clinics can then send us an email informing us. A pre-cooled LN 2 shipper is then sent to the clinic and the semen samples are flash frozen and the LN 2 shipper then returned to a SCSA diagnostic testing lab.
Some clinics send frozen semen samples in well insulated dry ice containers.
SCSA clinical data are then sent via a secure WEB site to the physician or patient that ordered the test.
There are five main populations of sperm identified by the SCSA® test:
the percentage of sperm with immature chromatin. HDS sperm exhibit less chromatin condensation, resulting in increased DNA stainability. Some studies have shown that if the %HDS is above 36%, no pregnancies have occurred (Menezo)
Our SCSA clinical report lists four statistical categories of fertility potential. These were derived from a comprehensive study of male fertility potential without medical intervention and have been confirmed in additional studies.
< 15% DFI = Excellent to good pregnancy outcomes (excluding female infertility factors)
> 15% to < 25% DFI = good to fair pregnancy outcomes
> 25% to < 40% DFI = fair to poor pregnancy outcomes
> 40% DFI = Very poor pregnancy outcomes
Furthermore, the finding shows a graphical representation of the %DFI (DNA Fragmentation Index) and the %HDS (high DNA stainability)
The graph shows that 76% of the measured sperm shows no DNA fragmentation and 24% shows medium-high DNA fragmentation (DFI= 24%). Pregnancy naturally or with the help of an IUI is possible.
Current data shows that the probability of a successful pregnancy is significantly lower when the proportion of sperm with fragmented DNA is > 25%. Nevertheless, a DFI > 25% does not rule out a normal pregnancy. The 25% threshold is a statistical threshold. Therefore, if a man has a constant DFI >25%, he will be placed in a statistical group of men who have been shown in clinical trials to take longer to achieve natural pregnancy and more IVF cycles. An increased rate of spontaneous abortions is associated with this or pregnancy does not occur at all.
The ovum is believed to be able to repair a certain amount of DNA damage.
Moderate to high levels of DNA fragmentation (DFI) found in the red fluorescent sperm likely exceed the egg’s DNA repair capacity.
Marcos Meseguer et al found that when oocytes from patients were taken, for every 10% increase in DFI, there was a 1.31 times increase in probability of not achieving pregnancy. Whereas, the same was not seen when donor oocytes were employed suggesting the higher capability of young oocytes to repair sperm DNA damage.
REPEATED IUI / IVF / ICSI FAILURE : The reason behind the unsuccessful attempts at IUI/IVF/ICSI can be identified and managed
REPEATED MISCARRAGE : An explanation for repeated spontaneous abortions (RPL, repeated miscarriage) could be found and guided
POOR EMBRYO DEVELOPMENT AND QUALITY : The reason behind poor embryo quality could be the fragmented sperm DNA which could be corrected before the next treatment
MEN AT RISK FOR HIGHER OXIDATIVE STRESS:INITIAL SCREENING TEST : Especially for couples who have been attempting pregnancy for a long time to identify the best management strategy
UNEXPLAINED INFERTILITY : Couples desiring children with all normal diagnostic tests
In cases of high DFI, we suggest:
When a risk factor is identified, after avoidance / correction of the risk factor a repeat SCSA test after 3 months is advised.
Yes. We offer a SCSA test at half the cost for controlling the effect of avoidance / correction of a risk factor.
Please find a few examples below:
After quitting smoking for 84 days
3 months after varicocele correction surgery :
3 months after stopping pesticide exposure:
After 24-48 hours of abstinence, the samples should be taken by masturbation into sterile plastic sample containers. After liquefaction of the sample, the sperm concentration, motility and morphology are determined.
Cryopreservation should take place no more than 1 hour after delivery, since the DFI can be falsely raised after a longer waiting period.
Flash-freezing: Freeze 0.25mL or more of liquefied native ejaculate in 1-2mL cryovials. Cryoprotectants is not required! (Please freeze 2 cryovials per patient)
Samples should be frozen as soon as possible after liquefaction to minimize exposure to air, which can cause damage caused by reactive oxygen species.Cryovials should be tightly capped, labeled with two identifiers (e.g. name and date of birth) and placed in an upright position directly into the pre-chilled cryo-container. The container must be closed quickly.
Note: Cryoprotectants are not needed since quick-frozen sperm give the clearest SCSA® data. This feature is unique to mammalian sperm cells due to the highly condensed, crystalline nature of the nuclei.
Note: If the sperm concentration is adequate, less than 0.5 mL (two 0.25 mL aliquots) of semen may be sufficient for the SCSA® test. To perform the test, we require two 0.20 mL aliquots with a concentration of >0.5 million/ml. Please contact our laboratory if you have questions pertaining to the volume necessary for a sample of a certain concentration.
We ship a pre-chilled dry shipper. Dry shippers store the liquid nitrogen in porous material and thus ensure the safe transport of your cryopreserved samples in the nitrogen gas phase.
The low temperature in the container can be maintained for 7 days. It is therefore important to send it back as soon as possible using the mailed/supplied shipping label.
Note: liquid nitrogen is adsorbed by special materials inside the shipping container. Even if they are turned upside down during transport, no liquid nitrogen will leak out. Dry shippers are not considered dangerous.
After receiving the sample(s) in our laboratory, we need 4 working days to create a report, which we will send to you.
The cost of the test is 295 Euro.
* Samples are shipped in a pre-cooled shipper that we can provide either at ambient or pre-chilled temperatures. The pre-cooled shipper will be sent to the centre where you have provided your sample for the centre to ship it to us.
The test at present is not covered by the insurance.
Please write us an email to contact@scsalab.com and after contacting you, we will send toy a dry shipper for the patient sample to be sent to us.