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

Hye Ran Kim; Jeong Hwan Shin; Woo Yeong Jung; Jeong Nyeo Lee

doi:10.3343/kjlm.2006.26.2.131

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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

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.

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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.

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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
No. case	Karyotype	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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