PMID- 16596261
OWN - NLM
STAT- MEDLINE
DCOM- 20061024
LR  - 20171116
IS  - 1107-3756 (Print)
IS  - 1107-3756 (Linking)
VI  - 17
IP  - 5
DP  - 2006 May
TI  - Quantitative evaluation of partial deletions of the DAZ gene cluster.
PG  - 785-9
AB  - Partial deletions of the DAZ gene cluster are thought to cause spermatogenesis 
      impairment. The presence of homologous copies of this gene in the Y chromosome 
      does not allow PCR to be used for the identification of this abnormality. Hence, 
      sequence family variants (SFV), following amplification of sY581, sY587 and sY586 
      and subsequent enzymatic digestion with Sau3A, DraI and TaqI, respectively, and 
      the dual fiber fluorescence in situ hybridization (FISH) have been used to this 
      aim. However, SFV is not always able to identify single DAZ gene copy deletions. 
      We report a quantitative real-time PCR application to evaluate partial deletions 
      of the DAZ gene cluster. To accomplish this, we designed a probe on exon 6 of the 
      DAZ gene which is repeated 3 times in DAZ1, once in DAZ2 and DAZ3 and twice in 
      DAZ4. Five normozoospermic healthy men (C1-C5) having 4 DAZ gene copies by SFV 
      were selected. Fiber-FISH confirmed this outcome in C1-C4, but not in C5 who had 
      an incomplete DAZ gene cluster. The men underwent then quantitative real-time PCR 
      and C1 was arbitrarily selected as calibrator for the calculation of the DAZ gene 
      signals because of the lowest variation in the threshold cycles. Real-time PCR 
      identified 7.2+/-0.05 signals in C2-C4 and 5.4+/-0.05 signals in C5. The overall 
      coefficient of variation was 1.4+/-0.2%. The loss of two signals in this subject 
      may relate to a deletion of both DAZ2 and DAZ3 or of DAZ4 gene. Since SFV showed 
      clearly the presence of DAZ2, it may be hypothesized that C5 lacks DAZ4. In 
      conclusion, these data suggested that quantitative real-time PCR seems to be an 
      effective and reproducible technique that can be used to study the DAZ gene 
      cluster. In addition, the probe chosen for this approach may give indication on 
      the DAZ gene copy deleted.
FAU - D'Amico, G Letizia
AU  - D'Amico GL
AD  - Section of Endocrinology, Andrology and Internal Medicine, University of Catania, 
      I-95123 Catania, Italy.
FAU - Di Benedetto, Daniela
AU  - Di Benedetto D
FAU - Pezzino, Franca M
AU  - Pezzino FM
FAU - Giuffrida, Vincenza
AU  - Giuffrida V
FAU - Libra, Massimo
AU  - Libra M
FAU - Fichera, Marco
AU  - Fichera M
FAU - Mauceri, Grazia
AU  - Mauceri G
FAU - Rappazzo, Giancarlo
AU  - Rappazzo G
FAU - D'Agata, Rosario
AU  - D'Agata R
FAU - Vicari, Enzo
AU  - Vicari E
FAU - Travali, Salvatore
AU  - Travali S
FAU - Calogero, Aldo E
AU  - Calogero AE
LA  - eng
PT  - Journal Article
PL  - Greece
TA  - Int J Mol Med
JT  - International journal of molecular medicine
JID - 9810955
RN  - 0 (DAZ1 protein, human)
RN  - 0 (Deleted in Azoospermia 1 Protein)
RN  - 0 (Genetic Markers)
RN  - 0 (RNA-Binding Proteins)
SB  - IM
MH  - Adult
MH  - *Chromosome Deletion
MH  - Chromosomes, Human, Y/*genetics
MH  - Deleted in Azoospermia 1 Protein
MH  - Genetic Markers/genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Male
MH  - Multigene Family/*genetics
MH  - Oligospermia/diagnosis/genetics
MH  - RNA-Binding Proteins/*genetics
MH  - Sensitivity and Specificity
MH  - Sequence Tagged Sites
EDAT- 2006/04/06 09:00
MHDA- 2006/10/25 09:00
CRDT- 2006/04/06 09:00
PHST- 2006/04/06 09:00 [pubmed]
PHST- 2006/10/25 09:00 [medline]
PHST- 2006/04/06 09:00 [entrez]
PST - ppublish
SO  - Int J Mol Med. 2006 May;17(5):785-9.