Reliable gender screening for human preimplantation embryos, using multiple DNA target-sequences

Levinson, Gene; Fields, R.; Harton, G.; Palmer, Frances T.; Maddalena, Anne; Fugger, Edward F.; Schulman, Joseph D.

doi:10.1093/oxfordjournals.humrep.a137846

Cited by 59 publications

(22 citation statements)

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“…Currently new methods are being introduced to further help couples avoid passing on 300 X-linked disorders [24]. The preimplantation technique [25,26] has been combined with new technologies such as dual fluorescence in situ hybridization and polymerase chain reaction. These methods help select female offspring to avoid X-linked disorders.…”

Section: Discussionmentioning

confidence: 99%

A Controlled Study for Gender Selection Using Swim-Up Separation

Khatamee

Horn²,

Weseley³

et al. 1999

Gynecol Obstet Invest

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Objective: To evaluate the success for gender selection using a sample of semen separated by a modified swim-up technique. Design: We retrospectively compared the gender outcome of two treatments (A and B) for either a male or female offspring with those who conceived spontaneously. Setting: Private practice of one author (M.A.K.). Patients, Participants: The treatment groups consisted of 52 total pregnancies for couples who conceived by the separation technique. Of these 52 participants, 15 desired a female offspring and were placed into treatment A and 37 desired a male offspring and were placed into treatment B. The control groups consisted of 162 women who were presented with initial consultation for gender selection and conceived spontaneously. Control group A consisted of 80 women who initially chose a female offspring, and control group B consisted of 82 participants who initially chose a male. Interventions: In treatment group A, one timed intrauterine insemination (IUI) was carried out with the bottom 0.5 ml of the separated semen on cycle days 12–14, when the follicle was 18–22 mm. Patients in this group were also administered clomiphene citrate and human chorionic gonadotropin. In treatment group B, one timed IUI was done with the top 0.5 ml of the separated semen, when the follicle was 18–22 mm. Main Outcome Measure: The gender outcome of the pregnancies of two treatment and control groups was evaluated based on the known desired gender. Results: The success rate for conceiving a female child after intervention (treatment group A) was 86.7% effective (p = 0.002) as compared to the control group A. Couples seeking a male child (treatment group B) were 89.2% effective (p = 0.0002) as compared to the control group B. Conclusions: This study reveals that the modified swim-up method with additional monitoring results in statistically significant gender preselection.

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Section: Discussionmentioning

confidence: 99%

A Controlled Study for Gender Selection Using Swim-Up Separation

Khatamee

Horn²,

Weseley³

et al. 1999

Gynecol Obstet Invest

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“…For indirect test, five new markers (FMR1-CA332P, FMR1-AC287P, FMR1-AC210P, FMR1-CA525D and FMR1-GT624D, see Supplementary Table 1 for sequences), were combined with FRAXAC1 12 and AMELY gene detection for sex determination 13 in a multiplex, single-round PCR test. A 47.5 μl reaction mix with primers (see Supplementary Table 1 For direct test, four new markers (FMR1-CA332P, FMR1-AC287P, FMR1-CA525D and FMR1-GT624D) were combined with normal CGG repeats detection 14 and AMELY sequence in a multiplex, single-round PCR test.…”

Section: Pcr Reactionmentioning

confidence: 99%

Improving preimplantation genetic diagnosis for Fragile X syndrome: two new powerful single-round multiplex indirect and direct tests

et al. 2015

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Fragile X syndrome (FraX) is caused by the expansion of an unstable CGG repeat located in the Fragile X mental retardation 1 gene (FMR1) gene. Preimplantation genetic diagnosis (PGD) can be proposed to couples at risk of transmitting the disease, that is, when the female carries a premutation or a full mutation. We describe two new single-cell, single-round multiplex PCR for indirect and direct diagnosis of FraX on biopsied embryos. These tests include five unpublished, highly heterozygous simple sequence repeats, and the co-amplification of non-expanded CGG repeats for the direct test. Heterozygosity of the new markers ranged from 69 to 81%. The mean rate of non-informative marker included in the tests was low (26% and 23% for the new indirect and direct tests, respectively). This strategy allows offering a PGD for FraX to 96% of couples requesting it in our centre. A conclusive genotype was obtained in all cells with a rate of cells presenting an allele dropout ranging from 17% for the indirect test to 26% for the direct test. The new indirect test was applied for eight PGD cycles: 32 embryos were analysed, 9 were transferred and 3 healthy babies were born. By multiplexing these highly informative markers, robustness of the diagnosis is improved and the loss of potentially healthy embryos (because they are non-diagnosed or misdiagnosed) is limited. This may increase the chances of success of couples requesting a PGD for FraX, in particular, when premature ovarian insufficiency in premutated women leads to a reduced number of embryos available for analysis.

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“…Also, the so called built-in control techniques consist of a single pair of primers that amplify a Y-chromosome specific sequence and a second byproduct. In bovine embryo sexing, only two techniques with a single pair of primers have been reported performing double amplification: amplification of a male-specific 178 base pairs (bp) fragment and an unidentified 145 bp by product common to male and female, which acts as an internal control (Hirayama et al, 2004;Kageyama et al, 2004), and amelogenin amplification (Levinson et al, 1992;Ennis & Gallagher, 1994). Amelogenin, as any other built-in control techniques, is theoretically advantageous over multiple primers and sequences amplification, as the use of the same primers to male and female sequence eliminates misdiagnosis errors associated to specific primer amplification efficiency (Levinson et al, 1992;Ennis & Gallagher, 1994;Pajares et al, 2007;Colley et al, 2008).…”

Section: Embryo Sexing By Pcrmentioning

confidence: 99%

Comparative study of PCR-sexing procedures using bovine embryos fertilized with sex-sorted spermatozoa

Trigal¹,

Gómez²,

Díez³

et al. 2012

Span. j. agric. res.

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Sex determination in bovine embryos is a useful tool in reproductive biotechnology. This work compares two techniques of embryo sexing by polymerase chain reaction (PCR). Embryos were produced in vitro with sex-sorted sperm and two techniques of DNA lysis were tested (proteinase K versus heat shock). Subsequently, halves of each lysed sample was amplified both by amelogenin and BRY4a/SAT1 primers. Male embryos treated by both digestion methods and amplified by BRY4a/SAT1 gave higher rates of false negatives. Amelogenin amplification failed with embryos previously digested by proteinase K. In contrast, the lysis method allowed obtaining the best accuracy in terms of sex verification when using amelogenin amplification.Additional key words: amelogenin; blastocyst; BRY4a/SAT1; sex technique. ResumenEstudio comparativo de dos métodos de sexaje por PCR en embriones bovinos producidos con semen sexado La determinación del sexo en embriones bovinos es una herramienta útil en las biotecnologías reproductivas. Este trabajo compara dos técnicas de determinación del sexo en embriones mediante la reacción en cadena de la polimerasa (PCR). Los embriones fueron producidos in vitro con semen sexado y se analizaron dos técnicas de lisis de ADN (proteinasa K y choque térmico). Posteriormente, la mitad de cada muestra lisada fue amplificada por la amelogenina y por los cebadores BRY4a/SAT1. Los embriones machos tratados por ambos métodos de digestión y amplificados por BRY4a/SAT1 dieron mayores tasas de falsos negativos. La amplificación de la amelogenina no dio resultados en embriones previamente digeridos por proteinasa K; sin embargo, el método de lisis por choque térmico junto con la amplificación por amelogenina permitió obtener la mayor precisión en términos de verificación de sexo.Palabras clave adicionales: amelogenina; blastocistos; BRY4a/SAT1; técnica de sexaje.

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Reliable gender screening for human preimplantation embryos, using multiple DNA target-sequences

Cited by 59 publications

References 0 publications

A Controlled Study for Gender Selection Using Swim-Up Separation

A Controlled Study for Gender Selection Using Swim-Up Separation

Improving preimplantation genetic diagnosis for Fragile X syndrome: two new powerful single-round multiplex indirect and direct tests

Comparative study of PCR-sexing procedures using bovine embryos fertilized with sex-sorted spermatozoa

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