1 Centre for Medical Genetics and 2 Centre for Reproductive Medicine, Dutch-speaking Brussels Free University (Vrije Universiteit Brussel), Brussels, Belgium.
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Abstract |
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Key words: genetic counselling/ICSI/karyotype/male and female infertility/prenatal diagnosis
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Introduction |
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ICSI is indeed a more invasive procedure than routine IVF, as one spermatozoon is injected through the oocyte membrane and fertilization can ensue from sperm which could never have been used previously in fertility treatment (Van Steirteghem et al., 1993). Two types of risk were mentioned by several authors that were either ICSI procedure-dependent or -independent (Patrizio, 1995
). In the latter case, the risk relates to the cause of infertility leading to the need to perform ICSI.
The risk related to the procedure itself may be due to: (i) the physical and/or biochemical disturbance of the ooplasm or of the meiotic spindle (leading to aneuploidization by errors of mitosis during early cleavage divisions) (Tesarik, 1995; Hewitson et al., 1999
; Wang et al., 2001a
); (ii) errors in selection of the injection site (due to variability of the location of the metaphase II spindle leading to the selection of a damaging injection site) (Hardarson et al., 2000
; Wang et al., 2001b
); (iii) the injection of biochemical contaminants; and (iv) the injection of foreign, sperm-associated exogenous DNA (Perry et al., 1999
; Chan et al., 2000
).
Among the ICSI procedure-independent problems are: (i) the microinjection of sperm carrying a chromosomal anomaly (such as an aneuploidy or structural defect); (ii) the transmission of a genetic defect [such as a Yq deletion or cystic fibrosis (CF) mutation] which is often the origin of male-factor infertility; (iii) male gamete structural defect; (iv) anomalies of sperm activating factors; (v) potential for incorporating sperm mitochondrial DNA; and (vi) female gamete anomalies (oocyte age-related).
Initial data on fetal karyotypes after ICSI reported by the authors group were not alarming, as no chromosomal abnormality in 43 prenatal tests on 56 consecutive ICSI fetuses were found (Bonduelle et al., 1994). Another group, however, reported a high incidence of chromosomal abnormalities (9/71) in a limited group of prenatal investigations selected for maternal age (Int Veld et al., 1995
; Van Opstal et al., 1997
). These data were not confirmed by a number of other reports on prenatal tests, mainly performed for maternal age risks (Testart et al., 1996
; Govaerts et al., 1998
; Loft et al., 1999
; Van Golde et al., 1999
; Wennerholm et al., 2000a
; Antoni and Hamori, 2001
; Van Steirteghem et al., 2002
), although reasons for concern remained as no data from systematic chromosomal investigation of all children (prenatally or post-natally) were available except for data from our own group (Liebaers et al., 1995
; Bonduelle et al., 1999
, 2002
) and recent data showing a significant increase in karyotype anomalies in children born after ICSI compared with the normal population (Aboulghar et al., 2001
)
In fact, very few groups have asked for an agreement for prenatal testing before starting ICSI treatment, although some have insisted on the importance of prenatal testing after ICSI treatment (Govaerts et al., 1995)
Data in the literature are in consequence, case reports or data from high-risk situations such as presence of parental chromosomal anomalies, maternal age risk or ultrasound anomalies detected during pregnancy, and do not provide a correct estimation of the ICSI-related risk for chromosomal anomalies.
The authors have continued to offer prenatal testing over a period of 11 years (19902001) to all parents attending the fertility clinic for assisted fertilization procedures and have collected results on 1586 foetuses. The aim was to determine whether ICSI with gametes of parents with normal karyotypes leads to a higher risk of chromosomal anomalies for the children and if so, whether these tend to be ICSI procedure-related or related to the types of gametes used.
When ICSI with non-ejaculated sperm, either epididymal or testicular, was introduced, emphasis was placed on the fact that the risk of chromosomal aberration in the sperm might be even higher in men with non-obstructive azoospermia (Van Assche et al., 1996). Data on this subgroup of ICSI children were also collected.
From the moment of the introduction of ICSI at the authors centre, the treatment involved an informed consent to a prospective follow-up study, involving genetic counselling, pregnancy data, prenatal diagnosis and child follow-up. Partial data on this cohort were published previously (Bonduelle et al., 1994, 1995a
,b
, 1996a
,b
, 1998
, 1999
, 2002
; ESHRE Task Force, 1998
). In these publications, the authors failed to find any increased risk of major congenital malformations as compared with the general population data, but an increased frequency of chromosomal aberrations, mostly sex-chromosomal aneuploidies, was identified (Bonduelle et al., 1994
, 1995a
,b
, 1996a
,b
, 1998
, 1999
, 2002
; Van Steirteghem et al., 2002
).
In the present study, attention was focused on the results of the prenatal testing, in a cohort of 2622 ICSI pregnancies established between 1990 and 2001. The fetal karyotypes were also analysed in relation to the origin and characteristics of the sperm used for ICSI. All pregnancies were obtained in a single centre at the Dutch-speaking Brussels Free University.
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Materials and methods |
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In this study, only ICSI pregnancies with transfer of fresh embryos were considered. Pregnancies obtained after the use of mixed ICSI-IVF procedures or after ICSI in combination with preimplantation genetic diagnosis (PGD) were not taken into account. From the 2622 ICSI pregnancies obtained using ejaculated, epididymal or testicular sperm, 1586 fetuses were tested prenatally; for 1469 of the tested fetuses, ICSI had been carried out with ejaculated sperm, for 31 with epididymal sperm, for 63 ICSI with testicular sperm, and for 23 with donor sperm. Epididymal or testicular sperm was used in a total of 94 samples from patients with either obstructive (n = 83) or non-obstructive azoospermia (n = 9) and from two patients without testis biopsy (Table I). In 63 cases, sperm which had been cryopreserved was used, while in two cases a mixed procedure with frozen and fresh sperm was used.
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Before starting ICSI, couples were asked to agree to participate in a prospective clinical follow-up study of the children. Couples were also asked to sign an informed consent, including agreement to genetic counselling and to prenatal karyotyping. Prior to ICSI, all couples were evaluated for possible genetic problems, and a parental karyotype was performed routinely. When pregnant, all couples were strongly counselled during the first years of the programme to have a prenatal diagnosis, first because of possible risk factors due to the introduction of the novel techniques of assisted fertilization, and second because the prenatal tests showing an apparently higher incidence of de-novo chromosomal anomalies of 1.2% (Bonduelle et al., 1996b). In the first 320 pregnancies, 70% of the fetuses were tested (Bonduelle et al., 1996a
); gradually, pregnant women could be informed more precisely about the risk of de-novo chromosomal anomalies in ICSI pregnancies of couples presenting with normal constitutional karyotypes, and sampling rates fell gradually after 1996 to under 50% (Bonduelle et al., 2002
). An appropriate strategy for prenatal testing was followed for those pregnant couples who had an a priori risk of transmitting a genetic disease to their offspring, such as in cases of inherited structural chromosomal aberrations. The pros and cons of the different types of prenatal diagnosis were discussed in detail at approximately 68 weeks gestation; amniocentesis (AC) was suggested for singleton pregnancies, while chorionic villus sampling (CVS) was proposed for multiple pregnancies (De Catte et al., 1996
, 2000
). Chromosome preparations were obtained from cultured amniocytes according to a modified technique (Verma and Babu, 1989
), leading to a mean number of 400550 chromosome bands. Chromosome preparations were obtained by means of a previously described technique from either non-cultured (Gibas et al., 1987
) or cultured chorionic villus cells (Yu et al., 1986
). If indicated, prenatal tests for other genetic diseases were planned. At either the follow-up examinations or at birth, a small number of children were karyotyped post-natally because during pregnancy the parents had wanted to avoid the risk of miscarriage related to the prenatal test procedures, or because there was an indication to do so based on the physical examination.
Statistical analysis
Statistical analysis was performed using the SAS statistical package version 6.12. Statistical tests were applied two-tailed at the 5% level of significance. (Since this is a safety study, no multiplicity correction was applied to the significance level; indeed, the type II risk in the analysis which is increased when applying a correction for multiplicity to the significance level is considered more important than the type I risk.) Comparisons of the treatment groups for discrete variables were performed using Fishers exact test. When the comparison was controlled for age, the CochranMantelHaenzel test for discrete variables was applied.
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Results |
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For 1586 ICSI fetuses, a final prenatal karyotype result was obtained by taking a total of 1619 samples. Of the tested fetuses, 902 were singletons and 684 multiples. Prenatal samples were taken by CVS for 710 fetuses, leading to a conclusive karyotype result on CVS for 698 fetuses, or to inconclusive results and four failures for the remaining 12. Of the four failed CVS tests, one was followed by an AC, while of the eight inconclusive tests seven were followed by an AC. Amniotic fluid samples were obtained via AC in 901 cases. Of these, 892 were first AC samples (of which eight were post CVS) and nine were repeat samples because of a failed or inconclusive first AC. All together, conclusive karyotype results were obtained for 888 fetuses after AC. One cord blood sample was taken in order to confirm a previous AC result (Table II).
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The mean maternal age in all women tested was 33.5 (median 33.5); the median age was 33.3 years in patients who underwent a CVS, and 33.7 years in those who choose an AC. Abnormal fetal karyotypes were found in 47 cases (36 singletons and 11 twins) out of 1586 prenatally tested ICSI fetuses; 25 anomalies were de novo (of these, 10 were sex chromosomal anomalies and 15 were autosomal [either numerical (n = 8) or structural (n = 7) anomalies]. Among 22 abnormal karyotypes inherited, 21 were balanced and one was unbalanced; in 17 of these 22 cases the inherited structural defects were transmitted through the father (Table III). Of the 47 fetuses with a chromosomal anomaly, 11 were abortedthree with a numerical sex chromosomal anomaly and eight with autosomal anomalies (seven numerical de novo and one inherited unbalanced). Furthermore, one twin pregnancy was lost due to miscarriage of two fetuses after a CVS procedure for a twin pregnancy had indicated the presence of an inherited balanced chromosomal anomaly, and three stillbirths occurred after 20 weeks of pregnancy with de-novo chromosomal anomalies (one with a sex numerical chromosomal anomaly, one with an autosomal numerical anomaly, and one with a complex structural anomaly). Ultimately, 31 children carrying a chromosomal anomaly were born without clinical abnormality at birth (Appendix I
). Details on the karyotype of the parents, age of the mother, multiplicity, prenatal test procedure and results, outcome of the pregnancy, physical examination at birth and semen source and origin are described in Appendix I
. The mothers of fetuses with a prenatal de-novo chromosomal anomaly had a mean age of 34.5 (range: 25.643.9) years, whereas in the total tested group the mean age was 33.5 years; details of the age distribution are given in Appendix I
. For 13 (2.2%) of the 588 fetuses tested for a maternal age risk, the result was a de-novo anomaly. For 12/988 (1.2%) of the fetuses of mothers aged <35 years, who were tested only because of the ISCI procedure, the result was a de-novo anomaly.
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Abnormal karyotypes were also found in 338 karyotyped children at birth (and not tested prenatally) (Table IV: Appendix II
). Of the de-novo anomalies, two were sex chromosomal (0.59%), three were autosomal numerical (0.88%), and two were autosomal structural (0.59%); six inherited balanced karyotypes were identified (1.78%).
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For morphology, 14% or <14% normal morphology was considered; missing data (396/1564) were considered as absent in a first analysis and as abnormal (<14% morphology) in a second analysis.
For motility, 50% motile sperm versus <50% was considered; missing data (510/1564) were considered absent in a first analysis and considered to be <50% motility (but were not possible to evaluate in a second analysis). A statistically significant difference was observed for sperm concentration of <20x106/ml, where a frequency of 2.1% chromosomal anomalies in the offspring was found, versus a frequency of 0.24% where sperm concentration was
20x106 /ml (Fishers Exact test, P = 0.006). For lower threshold values of sperm concentration (<1x106, <5x106, <10x106 and <15x106/ml), no statistical difference in the frequency of chromosomal anomalies was observed (Table V
).
The frequency of prenatal de-novo chromosomal abnormalities was 1.21% if the mother was aged 35 years, versus 2.26% if she was aged >35 years. The relationship between sperm concentration and chromosomal anomalies was the same for maternal ages
35 and >35 years [BreslowDay test, P = 0.168 (NS)].
No statistical difference was observed on the basis of sperm morphology for analyses 1 [Fishers exact test, P = 0.517 (NS)] and 2 [Fishers exact test, P = 1.0 (NS)].
A statistically significant difference was observed for 50% motile sperm cells versus <50% in analysis 1 (Fishers exact test, P = 0.027) and analysis 2 (Fishers exact test, P = 0.014).
All but one of the patients with an abnormal motility and a fetus with a karyotype anomaly also had an abnormal concentration of <20x106/ml (cross table between concentration and motility; Fishers exact test, P = 0.001).
All but three prenatal chromosomal anomalies were found in fetuses after ICSI procedures with ejaculated sperm (Table VI). Three chromosomal anomalies were found prenatally after the use of epididymal or testicular sperm in a total of 94 samples, 63 of which were from a testicular source. In these fetuses, two anomalies (among 51 tested fetuses) were found after the use of obstructive testicular sperm; one was a de-novo and one an inherited anomaly. One de-novo anomaly was also found in a still very limited number of fetuses tested after use of sperm from patients with a non-obstructive pathology (nine with a testis biopsy and two unclassified patients). Until now, no post-natal chromosomal anomalies have been found after the use of either epididymal or testicular sperm.
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Discussion |
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Abnormal fetal karyotypes were found in 3.0% (47/1586) of the prenatally tested ICSI fetuses: 1.6% were de-novo and 1.4% were inherited. Of the inherited anomalies, 17/22 were due to a chromosomal structural anomaly in the father. As described previously (Bonduelle et al., 1999), approximately 5% of the ICSI children were at increased a priori risk of chromosomal anomalies due to the chromosomal aberrations in their parents, most often the fathers, with either sex-chromosomal anomalies or structural anomalies. The figure of 1.4% [95% confidence interval (CI) 0.872.09%] inherited chromosomal anomalies (1.3% for balanced anomalies and 0.06% for unbalanced) was significantly higher (P < 0.001) than the expected anomalies in prenatal data of 0.290.37% (0.260.28% for balanced and 0.040.09% for unbalanced) [(Hook et al., 1984
) on 24951 fetuses; (Hook and Cross, 1987
) on 61000 fetuses]. This higher figure was a direct consequence of the higher number of structural (or numerical) chromosomal anomalies in the fathers presenting with severe male-factor infertility that is often present in the patient population needing ICSI (Van Assche et al., 1996
; Mau et al., 1997
; Meschede et al., 1998
; Scholtes et al., 1998
; Tuerlings et al., 1998
; Peschka et al., 1999
). For these parents a prenatal test was strongly recommended since, depending on their specific chromosomal structural anomaly, there is large range of risks of unbalanced gametes leading to a higher frequency of miscarriage, stillbirth and abnormal offspring at birth due to chromosomal imbalance (Van Assche et al., 1999
). Only one non-balanced karyotype was found, a trisomy 21 due to a Robertsonian translocation 14,21 (carried by the father) for which a termination was performed. All the other children (21/22) inherited exactly the same chromosomal structural anomaly as one of the parents and were phenotypically normal at birth. However, as described in the literaturebut still the subject of controversythere might also in this group be a slight increase in mental retardation and/or malformation, probably due to minor chromosomal imbalances, secondary to the structural anomaly (Fryns et al., 1986
). Follow-up at an older age might still reveal problems in this group.
It is recommended that chromosomal testing be performed in all parents prior to ICSI treatment, as the risk of finding a chromosomal anomaly in the peripheral lymphocytes of both male patients and also perhaps in their wives (Van Assche et al., 1996; Tuerlings et al., 1998
; Scholtes et al., 1998
) is higher. For those couples carrying a chromosomal anomaly, counselling and discussion is needed on their chances of having a successful treatment and the risk of an unbalanced offspring (Meshede et al., 1998; Scholtes et al., 1998
; Tuerlings et al., 1998
; Peschka et al., 1999
; Givens, 2000
; Schreurs et al., 2000
). Moreover, alternatives such as PGD for aneuploidy screening (Staessen et al., 1996
) or structural anomalies (Van Assche et al., 1999
) can be offered in a number of situations, and for some couples the chances of obtaining a normal pregnancy are so reduced that the option of donor gametes should be discussed. The figure of unbalanced gametes in male patients with reciprocal translocations is highly variable, leading to as few as 25% normal gametes in some cases checked by in-situ hybridization on individual sperm cells (Van Assche et al., 2001
). Parents should also be informed about the possible higher risk of infertility, mainly for their male children if a structural chromosomal anomaly is found which is known from literature data to be related to male infertility (Van Assche et al., 1996
).
De-novo aberrations were found in 1.6% of the tested ICSI children, which was significantly higher (P < 0.001) than in the general newborn population where 0.45% de-novo anomalies are described (Jacobs, 1992 on 56 952 newborns) (Table III). This higher figure was mostly due to a higher figure of sex chromosomal aberrations. The incidence of sex-chromosomal aberrations at the time of prenatal diagnosis is comparable with the incidence at birth, since these aberrations are not critical to survival (Schreinemachers et al., 1982
). The figure of 0.6% (95% CI 0.301.16%) of sex-chromosomal aberrations can thus be compared with the total newborn population, and is significantly different and approximately 3-fold higher than previously reported figures of 0.19% (P = 0.002; Jacobs et al., 1992
on 56 952 newborns), and of 0.23% (P = 0.006; Nielsen and Wohlert, 1991
on 34 910 newborns) found in an unselected newborn population. The same figure is also much higher than the calculated expected figures of 0.13% in live births (Hook, 1981
), and the value of 0.27% found in AC at a maternal age of 35 years (Ferguson-Smith 1983
; Ferguson-Smith and Yates, 1984
on 52 965 prenatal tests).
For data on autosomal anomalies however, it must be considered that a certain number of chromosomal numerical anomalies (e.g. trisomy 18) often end in spontaneous miscarriage, and that a percentage of 1.0% autosomal anomalies (both numerical and structural) may be overestimated in comparison with a newborn population (Schreinemachers et al., 1982).
When compared with literature data on prenatal diagnosis (AC) in the general population, the figure of 1.6% de-novo anomalies (sex chromosomal and autosomal) in ICSI pregnancies with a mean maternal age of 33.5 years is significantly higher than the 0.87% de-novo anomalies expected for a mean maternal age of 35 years (no data under the age of 35 years were collected) (Ferguson-Smith, 1983; Ferguson-Smith and Yates, 1984
on 52 965 ACs) or than the 0.45% de-novo anomalies calculated on the basis of previous reports (Hook, 1981
; Schreinemachers et al., 1982
) at a maternal age of 33.5 years. When only numerical autosomal anomalies were considered, a figure of 0.5% (95% CI 0.220.99%) was found for a mean maternal age of 33.5 years compared with 0.33% (Hook, 1981
; Schreinemachers et al., 1982
) for the same age, which was not significant (Table III
). The fact also had to be taken into account that, for 14 of the fetuses, prenatal data were collected on CVS, and that the prenatal figures on CVS material were higher than figures for AC, based on data which predicted an overall overestimation of 33% by comparing CVS and AC data (Hook et al., 1988
).
The type of sex chromosomal anomalies found included six non-mosaic anomalies (two each with 47,XXX, 47,XXY and 47,XYY) and four mosaic situations of the X chromosome. As it was not possible to check the neonatal karyotype on all these cases, these mosaicisms may have occurred either early in the first cell cycles or later in the placental tissues The frequency of mosaicism for sex chromosomes was 0.190.25%, and the total figure of mosaicism in the prenatal results was 0.250.31%, which compared well with the value reported by others (Hsu and Perlis, 1984) on 179 663 genetic amniocenteses in the general population with a maternal age risk (Hsu, 1996) (Table VII
). There is experimental evidence that epigenetic mechanisms are altered in this type of conception (Tesarik and Mendoza, 1996
), leading to a higher degree of aneuploidization due to errors of mitosis during the early cleavage divisions. This hypothesis of a higher risk of post-zygotic events as a consequence of the ICSI procedure leading to a higher figure for chromosomal mosaicism does not seem to be confirmed in this series of results (Int Veld et al., 1995
).
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Of the 25 tested ICSI fetuses with de-novo aberrations, 10 were aborted (seven for autosomal and three for sex-chromosomal numerical aberrations) after extensive counselling of the couple. Termination of sex-chromosomal anomalies is often subject to debate, since only minor physical problems may occur; mental retardation is not the rule, but learning difficulties, developmental problems and problems of social adaptation may occur (Linden et al., 1996; Meschede and Horst, 1997
; Ratcliffe, 1999
; Robinson et al., 1999). Frequently, an unduly pessimistic vision in the past has led to stigmatization of these patients and high abortion rates. When parents were counselled according to data obtained from long-term follow-up studies of cohorts of children with sex-chromosomal aneuploidies, approximately one-third of them chose to terminate the pregnancy (Linden et al., 1996
). In infertile couples, it can be expected that there will be even more reluctance to terminate such pregnancies. However, this was not reflected in the present patients, where three out of nine (33%) couples, when fully informed, opted for pregnancy termination for a fetus with a sex-chromosomal aneuploidy.
The higher rate of aneuploidy observed in the present series is most likely related to the higher aneuploidy rate in the sperm of the fathers, as several studies have described a higher percentage of gonosomal aneuploidy present in males with severe oligoasthenoteratozoospermia (OAT) (Bernardini et al., 1997; Downie et al., 1997
; Pang et al., 1999
; Vegetti et al., 2000
; Shi and Martin, 2001
). Indeed, a normal karyotype in the father does not exclude the possibility of germ cell aneuploidy, since an altered intratesticular environment not only damages spermatogenesis but may also disrupt the mechanisms controlling chromosomal segregation during meiosis (Calogero et al., 2001
). Another group (Storeng et al., 1998
) described sperm from men requiring ICSI as having a significantly higher proportion of XX- and XY-bearing sperm cells, while others (Calogero et al., 2001
) described sperm aneuploidy as being negatively correlated with sperm concentration, and particularly to the percentage of normal forms. Numbers of XY-bearing sperm were very low however, suggesting that chromosomal non-disjunction occurs mainly during the second meiotic division. We may consider our observations as being in line with these fluorescence in-situ hybridization (FISH) data, and conclude that the higher frequency of chromosomal aberrations in sperm from men with OAT is a risk factor in ICSI treatment and is in itself at the origin of the higher percentage of de-novo chromosomal aberrations observed. This might also explain the finding of a slightly higher number of de-novo structural aberrations, although as yet insufficient data have been reported to confirm this, apart from the high rate of de-novo translocations described in normal males during meiosis (Kurahashi et al., 2001).
Three chromosomal anomalies were found prenatally after the use of epididymal or testicular sperm in a total of 94 samples. Two were found in prenatal tests performed for fetuses after the use of testicular sperm with an obstructive pathology (on 52 tested fetuses); of these, one was a de-novo and one an inherited anomaly. One de-novo anomaly was found in an albeit very limited number of fetuses tested after use of sperm from patients with a non-obstructive pathology (11 patients). Although these values were high for de-novo anomalies (3.2%, or 2/63), the patient numbers were too small to draw valid conclusions. However, special attention should be paid in the future to this group of patients.
In support of the hypothesis of male-bearing chromosomal aneuploidies, it is possible to cite the literature data demonstrating the paternal origin of the prenatally detected chromosomal aberrations after ICSI (Van Opstal et al., 1997) (six out of six cases involving a sex chromosome abnormality were of paternal origin), and one observation in our own data on the paternal origin of the extra X chromosome in a 47,XXY karyotype (see Appendix I
). From data in the literature, it would be expected that half of the supernumerary X chromosomes would come from each of the parents (Harvey et al., 1990
).
The ICSI children with sex-chromosomal anomalies and de-novo structural chromosomal anomalies detected prenatally were all phenotypically normal at birth. It is known, however, that approximately 510% of the children with de-novo structural anomalies will have developmental problems that are not always apparent at birth (Fryns et al., 1986).
Comparison between the prenatally tested and non-tested group is important in order to determine whether any selection bias might have played a role in the higher incidence of karyotype anomalies observed in ICSI. Maternal age was slightly higher in the tested group, but as the maternal age was older in the entire ICSI cohort compared with the general population, the present data were compared to a similar age group in the general population. On the other hand, the most important indication for prenatal testing is the ICSI-related risk as counselled early in pregnancy. The prenatal test is generally planned at about 68 weeks gestation, and very few patients choose to be tested beyond this time. Some pregnancies were also tested for a positive triple test, detected around the 16th week of pregnancy, but none of the results of these prenatal tests was abnormal. Some prenatal tests were performed for higher risk for neural tube defect, and for fetal growth retardation or fetal anomalies detected during pregnancy, but again none of these prenatal tests was abnormal. All of the fetuses which showed an abnormal result were originally tested for reasons such as maternal age risk (13/26), a parental chromosomal anomaly (all of the inherited anomalies) or ICSI procedure (13/26) (see Appendix I). All tests carried out for other indications were normal; hence, it can be concluded that the number of abnormal results was not reduced due to any selection bias of opting for a prenatal diagnosis, or not.
Sperm characteristics may have been different in the tested versus not-tested group. An assessment of the sperm used in all cycles (tested and not-tested) revealed that all three semen parameters (sperm density, motility and morphology) were abnormal in 49.1% of cycles, two semen values were abnormal in 29.4% of cycles, one semen value was abnormal in 15.4% of cycles, and all three semen values were normal in 6.1% of cycles (Table VIII). Semen assessment of the fathers of fetuses for which a prenatal test was carried out revealed that all three semen parameters were abnormal in 51.0% of cycles, two semen values were abnormal in 35.8% of cycles, one semen value was abnormal in 10.3% of cycles, and all three semen values were normal in 2.9% of cycles (Table VIII
). In the prenatally tested group, sperm parameters were slightly abnormal more often, which may suggest that for some parents the choice of having a prenatal test or not may have been influenced by the sperm quality. On the other hand, 95.9% of the prenatally tested cycles and 97.1% of the total cycles were performed with abnormal sperm, and all of the prenatal abnormalities were found in cycles where abnormal sperm was used. It is considered therefore that the present data on prenatal testing in ICSI in this subset of patients provide a representative figure of expected anomalies in a patient population for which ICSI is used.
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The risk estimation of 1.6% for a de-novo anomaly after ICSI is similar to the age risk in a woman aged 40 years. In most European countries, this riskand even a much lower riskis acceptable for social security reimbursement of the costs of a prenatal diagnosis. One study in Denmark showed the costbenefit ratio to be in favour of prenatal testing for a maternal age of 35 years (Goldstein and Philip, 1990). Anomalies after ICSI do not always lead to termination of pregnancy, and in the present study only 10 terminations were performed in 1586 tested fetuses, producing a calculated risk of 0.6%. Nevertheless, such a value is still sufficiently high to be cost-effective, and especially when it offers parents the choice to undergo prenatal testing, or not (Botkin, 1990
; Druzin et al., 1993
).
In conclusion, the results of the present study indicate a higher risk for de-novo chromosomal anomalies that is mainly related to a higher level of sex-chromosomal anomalies, and is also partly related to a higher level of de-novo structural anomalies. In the light of these data on prenatal karyotypes, by taking into account various published data (Table IX), and by considering further observations that parents might require pregnancy interruption for certain conditions, we feel it necessary to continue offering prenatal diagnosis to all ICSI couples, and certainly to couples where the male partner has a sperm concentration <20x106/ml. However, couples should be informed of the risks of an abnormal result, of the risk linked to a prenatal procedure, and also be provided with information on the relatively benign character of certain de-novo structural or sex-chromosomal anomalies, in order for them to make any decision regarding prenatal testing.
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Acknowledgements |
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Notes |
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References |
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Submitted on March 28, 2002; accepted on June 19, 2002.