Molecular Human Reproduction, Vol. 8, No. 6, 589-595,
June 2002
© 2002 European Society of Human Reproduction and Embryology
Reproductive genetics |
Genetic follow-up of male offspring born by ICSI, using a multiplex fluorescent PCR-based test for Yq deletions
1 Monash Institute of Reproduction and Development, Monash University, Monash Medical Centre, 2 Prince Henry's Institute of Medical Research, Monash Medical Centre and 3 Monash IVF Pty Ltd, Clayton Rd, Clayton, Victoria 3168, Australia
De-novo deletions involving AZFa, b, c and d are one of the most common chromosomal aberrations in man resulting in defective spermatogenesis and male infertility. Currently, Yq deletion screening involves either single or multiplex PCR using Y-specific sequence tagged site markers and the subsequent analysis of the amplification products on ethidium bromide-stained agarose gels. To improve the practicality of routine and high throughput Yq testing, we have developed a more sensitive multiplex fluorescent (FL)–PCR screening system using genomic DNA extracted from cheek buccal cells as a readily available PCR template. For genetic follow-up studies of ICSI-conceived children, we also developed a DNA fingerprinting system based on the co-amplification of four highly polymorphic markers to validate family samples and detect any potential extraneous DNA contamination that could cause a misdiagnosis. Multiplex FL–PCR analysis of buccal cell DNA from two infertile men who conceived three sons by ICSI demonstrated that their Yq deletions were vertically transmitted. Fine mapping with additional Yq markers revealed identical deletion endpoints involving the loss of AZFdc sequences. This firstly indicates that the extent of the Yq deletion was unchanged on ICSI transmission and secondly supports the view that AZFdc deletions may arise by a common de-novo event. Analysis of paternal, maternal and sibling DNA fingerprints showed the co-inheritance of parental alleles by each male child and confirmed the expected relationship between each family member. The application of these new FL–PCR based screening tests in genetic follow-up studies will assist in confirming transmission of specific genetic defects to male offspring conceived by ICSI and provide a basis for genetic counselling and potential treatment options as these boys approach sexual maturity.
DNA fingerprinting/ICSI/male infertility/multiplex FL-PCR/Yq deletions
4 To whom correspondence should be addressed at: MIRD, level 3, 27–31 Wright St, Clayton, Victoria 3168, Australia.E-mail: mandy.katz{at}med.monash.edu.au
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. A. Ward Intracytoplasmic Sperm Injection Effects in Infertile azh Mutant Mice Biol Reprod, July 1, 2005; 73(1): 193 - 200. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Devroey and A. Van Steirteghem A review of ten years experience of ICSI Hum. Reprod. Update, January 1, 2004; 10(1): 19 - 28. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Joffe Infertility and environmental pollutants Br. Med. Bull., December 1, 2003; 68(1): 47 - 70. [Abstract] [Full Text] [PDF]
|
|