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Journal of Assisted Reproduction and Genetics

Published in:

01-01-2012 | Genetics

Preimplantation genetic diagnosis of alpha-thalassemia^-SEA using novel multiplex fluorescent PCR

Authors: Wirawit Piyamongkol, Teraporn Vutyavanich, Torpong Sanguansermsri

Published in: Journal of Assisted Reproduction and Genetics | Issue 1/2012

Abstract

Purpose

Preimplantation genetic diagnosis (PGD) is an alternative to prenatal diagnosis (PND) giving couples at risk a chance to start a pregnancy with a disease-free baby. This study aimed to develop a new PGD protocol for alpha-thalassemia^-SEA mutation, the commonest Mendelian disorder.

Patients and methods

Multiplex fluorescent PCR was employed for mutation, contamination and linkage analysis. A couple experienced termination of pregnancy following positive PND decided to join the project.

Results

Novel primers for alpha-thalassemia^-SEA mutation amplifying 5 DNA fragments were developed. Two PGD cycles were performed, resulting in an un-affected baby. PND confirmed the heterozygous result. From 24 embryos, 87.5% of affected genotype were of best quality compared to 0% and 18.2% of those with normal and heterozygous, respectively.

Conclusions

A novel PCR protocol for the common alpha-thalassemia^-SEA mutation is reported. This test should be widely applicable. Interestingly, a potential effect of alpha-thalassemia^-SEA mutation on preimplantation embryonic development was noticed.

Weatherall DJ. Disorders of the synthesis or function of haemoglobin. In: Weatherall DJ, Ledingham JGG, Warrell DA, editors. Oxford textbook of medicine. 3rd ed. Oxford: Oxford University Press; 1996. p. 3500–20.

Handyside AH, Pattinson JK, Penketh RJ, Delhanty JD, Winston RM, Tuddenham EG. Biopsy of human preimplantation embryos and sexing by DNA amplification. Lancet. 1989;1(8634):347–9.PubMedCrossRef

Staessen C, Camus M, Bollen N, Devroey P, Van Steirteghem AC. The relationship between embryo quality and the occurrence of multiple pregnancies. Fertil Steril. 1992;57(3):626–30.PubMed

El-Hashemite N, Delhanty JD. A technique for eliminating allele specific amplification failure during DNA amplification of heterozygous cells for preimplantation diagnosis. Mol Hum Reprod. 1997;3(11):975–8.PubMedCrossRef

Piyamongkol W, Harper JC, Sherlock JK, Doshi A, Serhal PF, Delhanty JD, et al. A successful strategy for preimplantation genetic diagnosis of myotonic dystrophy using multiplex fluorescent PCR. Prenat Diagn. 2001;21(3):223–32.PubMedCrossRef

Piyamongkol W, Vutyavanich T, Piyamongkol S, Wells D, Kunaviktikul C, Tongsong T, et al. A successful strategy for Preimplantation Genetic Diagnosis of beta-thalassemia and simultaneous detection of Down’s syndrome using multiplex fluorescent PCR. J Med Assoc Thai. 2006;89(7):918–27.PubMed

Hattori M, Yoshioka K, Sakaki Y. High-sensitive fluorescent DNA sequencing and its application for detection and mass-screening of point mutations. Electrophoresis. 1992;13(8):560–5.PubMedCrossRef

Tongsong T, Wanapirak C, Sirivatanapa P, Sanguansermsri T, Sirichotiyakul S, Piyamongkol W, et al. Prenatal control of severe thalassaemia: Chiang Mai strategy. Prenat Diagn. 2000;20(3):229–34.PubMedCrossRef

Wells D, Delhanty JD. Preimplantation genetic diagnosis: applications for molecular medicine. Mol Med Today (Trends Mol Med). 2001;7(1):23–30.

10.

Ko TM, Tseng LH, Hsieh FJ, Hsu PM, Lee TY. Carrier detection and prenatal diagnosis of alpha-thalassemia of Southeast Asian deletion by polymerase chain reaction. Hum Genet. 1992;88(3):245–8.PubMedCrossRef

11.

Piyamongkol W, Bermudez MG, Harper JC, Wells D. Detailed investigation of factors influencing amplification efficiency and allele drop-out in single cell PCR: implications for preimplantation genetic diagnosis. Mol Hum Reprod. 2003;9(7):411–20.PubMedCrossRef

Title: Preimplantation genetic diagnosis of alpha-thalassemia-SEA using novel multiplex fluorescent PCR
Authors: Wirawit Piyamongkol
Teraporn Vutyavanich
Torpong Sanguansermsri
Publication date: 01-01-2012
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 1/2012
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-011-9641-6

At a glance: The STEP trials

Springer Medicine

Preimplantation genetic diagnosis of alpha-thalassemia^-SEA using novel multiplex fluorescent PCR

Abstract

Purpose

Patients and methods

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Patients and methods

Results

Conclusions

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