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Kim, Shin, Jung, and Lee: Identification of Y-chromosome by Molecular Analysis in Patients with Turner Syndrome
Original Article

Korean J Leg Med 2006;26(2):131-136.

Published online: 10 February 2006

DOI: https://doi.org/10.3343/kjlm.2006.26.2.131

Identification of Y-chromosome by Molecular Analysis in Patients with Turner Syndrome

Hye Ran Kim, M.D.1, Jeong Hwan Shin, M.D.1, 2, Woo Yeong Jung, M.D.3, Jeong Nyeo Lee, M.D.1, 2

1Department of Laboratory Medicine, Busan Paik Hospital, College of Medicine, Inje University, Busan, Korea

2Department of Pediatrics, Busan Paik Hospital, College of Medicine, Inje University, Busan, Korea

3Department of Paik Institute for Clinical Research, Busan Paik Hospital, College of Medicine, Inje University, Busan, Korea

Received 20 January 2006       Revised 21 February 2006       Accepted 15 March 2006

Copyright © 2006 The Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

It is known that the Y chromosome or Y-specific sequence is present in about 6% of Turner syndrome (TS) patients and that it predisposes them to gonadoblastoma formation with an estimated risk of 15–25%. In this study, we performed a polymerase chain reaction (PCR) in 32 patients with TS to detect Y-specific sequence. The results were compared with those obtained by the fluorescence in situ hybridaization (FISH) method.

Methods

Cytogenetic analysis was performed by phytohaemagglutinin (PHA)-stimulated peripheral lymphocyte cultures, using G-banding. DNA was extracted from peripheral blood for PCR. Seven different sets of oligonucleotide primers, sex determining region Y (SRY), zinc finger gene on the Y chromosome (ZFY), testis specific protein Y (TSPY), DYZ3, DYF49S1, RNA binding motif protein (RBM), and DYZ1, spanning on centromeres and short and long arms of the Y chromosome were used for PCR. FISH was carried out using X and Y chromosome enumeration probe for Xp11.1-q11.1 (DXZ1 locus) and Yp11.1-q11.1 (DYZ3 locus), respectively.

Results

Among 32 patients with TS, four (12.5%) were positive for Y specific sequence by PCR. Of these, two patients were detected previously by a cytogenetic analysis: 45,X/47,XYY and 45,X/46,XY. Only one Y specific sequence, DYZ3, was detected by PCR in the other two patients without cytogenetically obvious Y chromosome. Y signal was not detected by FISH for the last two patients.

Conclusions

It may be reasonable to consider using a PCR method to screen for Y-specific sequences in all patients with TS. Even though we did not demonstrate Y-signal by FISH in patients with PCR positive and cytogenetically no obvious Y chromosome, FISH may be another useful method in TS patient, and futher investigation is nessessary.

Keywords: Turner syndrome, Y-chromosome, Gonadoblastoma, PCR, FISH

References

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Fig. 1.
Multiplex PCR amplification of the DYZ3 gene (M, marker; C, negative control; P, positive control; lanes 1–4, Cases 1–4 was described in Table 3).
Abbreviation: PCR, polymerase chain reaction.
kjlm-26-131f1.tif
Table 1.
Primers and product for PCR amplication
Markers Intervals Primers Product size Reference
SRY 1A1A 5′-CAGTGTAAACGGGAGAAAACAGT-3′ 270 bp [6]
    5′-CTTCCGACGAGGTCGATACTTATA-3′    
ZFY 1A2 5′-CGAATTCATACCGGCGAGAAGCCATACC-3′ 735 bp [7]
    5′-AAAGCTTGTAGACACATCGTTAGGG-3′    
TSPY 3C 5′-CCAGATGTCAGCCCTGATCA-3′ 450 bp [8]
    5′-GCTCCATCATATTCAACTCAAT-3′    
DYZ3 4B 5′-TCCTTTTCCACAATAGACGTCA-3′ 170 bp [8]
    5′-GGAAGTATCTTCCCTTAAAAGCTA-3′    
DYF49S1 6B 5′-CACATGAAGCACTGGAACTG-3′ 170 bp [8]
    5′-AGGGCCTGAGTCTCCAGG-3′    
RBM 3 5′-CTCGGATGTCTTATGGTGGAA-3′ 474 bp [8]
    5′-GCATCAACAAGTATGAAATTACT-3′    
DYZ1 7 5′-TACGGGTCTCGAATGGAATA-3′ 236 bp [8]
    5′-TCATTGCATTCCTTTCCATT-3′    

Abbreviations: PCR, polymerase chain reaction; SRY, sex determining region Y; ZFY, zinc finger gene on the Y chromosome; TSPY, testis specific protein Y; RBM, RNA binding motif protein.

Table 2.
Various karyotypes of 32 patients with Turner syndrome
Karyotype Number of patient (%)
45,X 16 (50.0)
45,X/46,X,i(Xq) 4 (12.5)
46,X,i(Xq) 3 (9.4)
45,X/47,XXX 2 (6.2)
45,X/47,XXX/46,XX 2 (6.2)
45,X/46,XX 1 (3.2)
45,X/47,XYY 1 (3.2)
45,X/46,XY 1 (3.2)
45,X/46,X,+mar/46,XX 1 (3.2)
Total 32
Table 3.
Summary of positive results of PCR and FISH analyses for presence of Y-chromosome material
No. case Karyotype PCR
 FISH
SRY DYZ3 DYF49S RBM DYZ1 ZFY TSPY DXZ1 DYZ3
1 47,XYY[36]/45,X[14] + + + + + + + + +
2 45,X[37]/46,XY[13] + + + + + + − + +
3 45,X[92]/46,X,+mar[3]/46,XX[5] − + − − − − − + −
4 45,X[61]/47,XXX[35]/46,XX[4] − + − − − − − + −

Abbreviations: PCR, polymerase chain reaction; FISH, fluorescence in situ hybridization; SRY, sex determining region Y; ZFY, zinc finger gene on the Y chromosome; TSPY, testis specific protein Y; RBM, RNA binding motif protein.

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