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Evaluation of Ovarian Reserve Following Tubal Sterilization Using Anti-Müllerian Hormone
in misdiagnosis at a rate of 1-2% or in false positives when no maternal haplotype is available either from polar bodies or grandparent samples. Efficiency and reliability can be improved by using a SNP microarray and informatics to phase the parent homologs without grandparent samples and to infer recombination sites. OBJECTIVE(S): To evaluate the clinical benefit of a novel informaticsbased technology that screens for single gene disorders in parallel with 24chromosome aneuploidy in at-risk couples. MATERIALS AND METHOD(S): Seventeen couples at risk of having a child affected with a single gene disorder were referred for PGD testing during in vitro fertilization. The couples were carriers for the following: Gaucher’s disease, cystic fibrosis, Walker-Warburg syndrome, Duchenne muscular dystrophy, Tay Sachs, Alport syndrome, Gardner’s syndrome, Fragile X, retinoblastoma, Fabry disease, Type 1 neutrofibromatosis syndrome and X-linked adrenoleukodystrophy. Following Day 3 biopsy, single blastomeres were shipped on dry ice to a laboratory for testing with the informatics-based technology. RESULT(S): Of the 17 couples, 14 had a transfer on Day 5/6 with embryos that tested as unaffected for the target genetic locus and were also euploid across all 24 chromosomes. Of the 14 couples, 11 (79%), achieved chemical pregnancy. On average, 40 informative markers are found in the region of the genetic locus of interest, and average confidence of genotype calls is above 99.9%. Amniocentesis or CVS was so far performed in three couples with results confirming the initial Day 3 biopsy. CONCLUSION(S): Reliable single gene testing in parallel with 24-chromosome aneuploidy screening can be performed with informatics enhanced SNP microarrays using a single cell. Preliminary results show highly accurate single gene calls and 79% chemical pregnancy rates. SUPPORT: None.
O-6 Implantation and Miscarriage Rates Following Array CGH Analysis at the Cleavage and Blastocyst Stages. G. Harton, M. Surrey, J. Grifo, B. Kaplan, S. Munne. OBJECTIVE(S): To assess the effect of maternal age on implantation after PGD with Comparative Genomic Hybridization (CGH) or microarray CGH (aCGH) after biopsy at day-3 (clevage) or day-5 (blastocyst) of embryo development. DESIGN: Retrospective multi-center comparative study. MATERIAL AND METHOD(S): PGD Patients from various IVF clinics had embryos biopsied on day-3 or 5. Analysis on day-3 involved 1-cell biopsy, aCGH and transfer of euploid embryos on day-5. Analysis of blastocysts involved trophectoderm biopsy on day-5 or -6 followed by CGH, vitrification and transfer of euploid embryos in a later cycle. RESULT(S):
Age group
Implant (sac)
Sacs lost
Day 3 Day 5
30–34 30–34
Day 3 Day 5
35–39 35–39
Day 3 Day 5
40–42 40–42
51.4 (70/136) 85.7 (24/28) P<0.005 38.2 (63/165) 63.8 (23/36) P<0.01 31.0 (40/121) 77.7 (28/36) P<0.001
7.1 (5/70) 8.3 (2/24) NS 9.5 (6/63) 4.3 (1/23) NS 15.0 (6/40) 7.1 (2/28) NS
O-7 Evaluation of Ovarian Reserve Following Tubal Sterilization Using Anti-M€ ullerian Hormone. M. A. Grant, G. E. Chow, R. O. Burney, Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Madigan Army Medical Center, Tacoma, Washington. BACKGROUND: Tubal sterilization is among the most frequently used methods of contraception in the United States. Depending on the type of tubal sterilization procedure, the mesosalpingeal vascular arcade, a source of collateral ovarian blood flow, is variably interrupted. In women presenting for desired fertility after previous tubal sterilization, ovarian reserve is a critical determinant as to whether tubal anastamosis or in vitro fertilization represents the best treatment option for the couple. Antimullerian hormone (AMH) is an emerging marker for the primary measurement of ovarian reserve. OBJECTIVE(S): To assess the impact of tubal sterilization history on ovarian reserve. MATERIALS AND METHOD(S): Ovarian reserve metrics to include AMH were available for 300 patients presenting for infertility from March 2008 to April 2010 at a single tertiary care center. Of these, 50 patients were regularly menstruating women with a history of tubal sterilization desiring fertility. Healthy women from couples diagnosed with male factor infertility only (n ¼ 33) comprised the control group. Women diagnosed with PCOS were excluded from analysis. AMH, antral follicle count (AFC), basal follicle stimulating hormone (FSH), FSH/LH ratio, and estradiol were compared between groups. RESULT(S): There were no significant differences in mean age or body mass index between the two groups. Mean basal FSH, FSH/LH ratio and estradiol levels were comparable. However, the total AFC (12.87 vs. 17.27; P¼.0003) and serum AMH (1.74 vs. 2.71; P¼.001) were significantly reduced in women with a history of tubal sterilization compared to women with normal tubal status. When stratified by type of tubal sterilization, methods with greater potential for disruption of collateral ovarian blood supply (i.e. electrocautery) were associated with lower AMH levels.
Aneup rate 50.5 40.4 P<0.005 65.1 49.2 P<0.005 77.8 61.1 P<0.001
CONCLUSION(S): There were fewer abnormal embryos when biopsied at the blastocyst stage compared with embryos biopsied at the cleavage stage, suggesting selection against aneuploidy during extended culture. However, blastocyst aneuploidy rates remained high and increased with advancing maternal age. Interestingly, implantation rates for euploid blastocysts were not significantly different across all age groups, however, for day-3 biopsy, implantation rates declined with age. Since both techniques
FERTILITY & STERILITYÒ
detect the same abnormalities, the decrease in implantation with age following biopsy at day-3 is unlikely to be a screening problem and suggests the involvement of non-chromosomal factors. The impact of biopsy on embryos at the clevage stage versus the blastocyst stage is largely unknown. An age-related uterine receptivity problem may be magnified after controlled ovarian stimulation and alleviated by transfer of diagnosed blastocysts in unstimulated cycles. There are several limitations of this retrospective analysis; however the findings are intriguing enough to warrant further study.
CONCLUSION(S): To our knowledge, this is the first study to evaluate the impact of tubal sterilization on ovarian reserve using serum AMH, a marker directly related to the primary follicle pool. Our results suggest an association between tubal sterilization and reduced ovarian reserve. Further study is necessary to delineate a possible relationship between type of tubal sterilization and ovarian reserve parameters. SUPPORT: Departmental funds.
S7
O-6 Implantation and Miscarriage Rates Following Array CGH Analysis at the Cleavage and Blastocyst Stages. G. Harton, M. Surrey, J. Grifo, B. Kaplan, S. Munne. OBJECTIVE(S): To assess the effect of maternal age on implantation after PGD with Comparative Genomic Hybridization (CGH) or microarray CGH (aCGH) after biopsy at day-3 (clevage) or day-5 (blastocyst) of embryo development. DESIGN: Retrospective multi-center comparative study. MATERIAL AND METHOD(S): PGD Patients from various IVF clinics had embryos biopsied on day-3 or 5. Analysis on day-3 involved 1-cell biopsy, aCGH and transfer of euploid embryos on day-5. Analysis of blastocysts involved trophectoderm biopsy on day-5 or -6 followed by CGH, vitrification and transfer of euploid embryos in a later cycle. RESULT(S):
Age group
Implant (sac)
Sacs lost
Day 3 Day 5
30–34 30–34
Day 3 Day 5
35–39 35–39
Day 3 Day 5
40–42 40–42
51.4 (70/136) 85.7 (24/28) P<0.005 38.2 (63/165) 63.8 (23/36) P<0.01 31.0 (40/121) 77.7 (28/36) P<0.001
7.1 (5/70) 8.3 (2/24) NS 9.5 (6/63) 4.3 (1/23) NS 15.0 (6/40) 7.1 (2/28) NS
O-7 Evaluation of Ovarian Reserve Following Tubal Sterilization Using Anti-M€ ullerian Hormone. M. A. Grant, G. E. Chow, R. O. Burney, Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Madigan Army Medical Center, Tacoma, Washington. BACKGROUND: Tubal sterilization is among the most frequently used methods of contraception in the United States. Depending on the type of tubal sterilization procedure, the mesosalpingeal vascular arcade, a source of collateral ovarian blood flow, is variably interrupted. In women presenting for desired fertility after previous tubal sterilization, ovarian reserve is a critical determinant as to whether tubal anastamosis or in vitro fertilization represents the best treatment option for the couple. Antimullerian hormone (AMH) is an emerging marker for the primary measurement of ovarian reserve. OBJECTIVE(S): To assess the impact of tubal sterilization history on ovarian reserve. MATERIALS AND METHOD(S): Ovarian reserve metrics to include AMH were available for 300 patients presenting for infertility from March 2008 to April 2010 at a single tertiary care center. Of these, 50 patients were regularly menstruating women with a history of tubal sterilization desiring fertility. Healthy women from couples diagnosed with male factor infertility only (n ¼ 33) comprised the control group. Women diagnosed with PCOS were excluded from analysis. AMH, antral follicle count (AFC), basal follicle stimulating hormone (FSH), FSH/LH ratio, and estradiol were compared between groups. RESULT(S): There were no significant differences in mean age or body mass index between the two groups. Mean basal FSH, FSH/LH ratio and estradiol levels were comparable. However, the total AFC (12.87 vs. 17.27; P¼.0003) and serum AMH (1.74 vs. 2.71; P¼.001) were significantly reduced in women with a history of tubal sterilization compared to women with normal tubal status. When stratified by type of tubal sterilization, methods with greater potential for disruption of collateral ovarian blood supply (i.e. electrocautery) were associated with lower AMH levels.
Aneup rate 50.5 40.4 P<0.005 65.1 49.2 P<0.005 77.8 61.1 P<0.001
CONCLUSION(S): There were fewer abnormal embryos when biopsied at the blastocyst stage compared with embryos biopsied at the cleavage stage, suggesting selection against aneuploidy during extended culture. However, blastocyst aneuploidy rates remained high and increased with advancing maternal age. Interestingly, implantation rates for euploid blastocysts were not significantly different across all age groups, however, for day-3 biopsy, implantation rates declined with age. Since both techniques
FERTILITY & STERILITYÒ
detect the same abnormalities, the decrease in implantation with age following biopsy at day-3 is unlikely to be a screening problem and suggests the involvement of non-chromosomal factors. The impact of biopsy on embryos at the clevage stage versus the blastocyst stage is largely unknown. An age-related uterine receptivity problem may be magnified after controlled ovarian stimulation and alleviated by transfer of diagnosed blastocysts in unstimulated cycles. There are several limitations of this retrospective analysis; however the findings are intriguing enough to warrant further study.
CONCLUSION(S): To our knowledge, this is the first study to evaluate the impact of tubal sterilization on ovarian reserve using serum AMH, a marker directly related to the primary follicle pool. Our results suggest an association between tubal sterilization and reduced ovarian reserve. Further study is necessary to delineate a possible relationship between type of tubal sterilization and ovarian reserve parameters. SUPPORT: Departmental funds.
S7