Validation and comparison of two quantitative real-time PCR assays for direct detection of DMD/BMD carriers

被引：22

作者：

Nosaeid, Mina Hayat ^{[1
]}

Mahdian, Reza ^{[1
]}

Jamali, Somayeh ^{[1
]}

Maryami, Fereshteh ^{[1
]}

Babashah, Sadegh ^{[1
,2
]}

Maryami, Fahimeh ^{[1
]}

Karimipoor, Morteza ^{[1
]}

Zeinali, Sirous ^{[1
,2
]}

机构：

[1] Pasteur Inst Iran, Dept Mol Med, Biotechnol Res Ctr, Tehran, Iran

[2] Kawsar Human Genet Res Ctr, Tehran, Iran

来源：

CLINICAL BIOCHEMISTRY | 2009年 / 42卷 / 12期

关键词：

Duchenne muscular dystrophy; Quantitative real-time PCR; Deletion mutation; Carrier detection; DUCHENNE MUSCULAR-DYSTROPHY; POLYMERASE-CHAIN-REACTION; GENE DELETIONS; RELATIVE QUANTIFICATION; PROBE-AMPLIFICATION; IDENTIFICATION; REARRANGEMENTS; DUPLICATIONS; DIAGNOSIS; LOCUS;

D O I：

10.1016/j.clinbiochem.2009.04.016

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

100118 [医学信息学]; 100208 [临床检验诊断学];

摘要：

Objectives: To develop a robust and reliable assay for direct identification of female carriers of deletions in the dystrophin gene. Design and methods: We compared two quantitative real-time PCR approaches for the detection of the deletions of exons 4, 17, 47, and 50 in DMD/BMD carriers. One hundred and ten individuals from 26 unrelated families, including 8 large pedigrees characterized by having at least two DMD affected males, were studied. Carrier status of the subjects was also evaluated by MLPA. Results: The results showed the gene dosage ratio of 0.99 +/- 0.14 and 1.09 +/- 0.19 for normal individuals and 0.48 +/- 0.06 and 0.50 +/- 0.10 for carriers in SYBR green and TaqMan probe assays, respectively. Carrier status was accurately attributed in 100% of cases and confirmed by MLPA. Conclusion: Quantitative real-time PCR can be used as a direct method for carrier detection in female relatives of DMD patients with known deletions. The results are comparable to the MLPA data. (C) 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

引用

页码：1291 / 1299

页数：9

共 25 条

[1]

BEGGS AH, 1990, HUM GENET, V86, P45

[2]

Calvano S, 1997, CLIN GENET, V52, P17

[3]

DELETION SCREENING OF THE DUCHENNE MUSCULAR-DYSTROPHY LOCUS VIA MULTIPLEX DNA AMPLIFICATION [J].

CHAMBERLAIN, JS ;

GIBBS, RA ;

RANIER, JE ;

NGUYEN, PN ;

CASKEY, CT .

NUCLEIC ACIDS RESEARCH, 1988, 16 (23) :11141-11156

[4]

Quality control in molecular genetic testing [J].

Dequeker, E ;

Ramsden, S ;

Grody, WW ;

Stenzel, TT ;

Barton, DE .

NATURE REVIEWS GENETICS, 2001, 2 (09) :717-723

[5]

Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA) [J].

Gatta, V ;

Scarciolla, O ;

Gaspari, AR ;

Palka, C ;

De Angelis, MV ;

Di Muzio, A ;

Guanciali-Franchi, P ;

Calabrese, G ;

Uncini, A ;

Stuppia, L .

HUMAN GENETICS, 2005, 117 (01) :92-98

[6]

HU XY, 1990, AM J HUM GENET, V46, P682

[7]

Rapid identification of female carriers of DMD/BMD by quantitative real-time PCR [J].

Joncourt, F ;

Neuhaus, B ;

Jostarndt-Foegen, K ;

Kleinle, S ;

Steiner, B ;

Gallati, S .

HUMAN MUTATION, 2004, 23 (04) :385-391

[8]

Kubista Mikael, 2006, Molecular Aspects of Medicine, V27, P95, DOI 10.1016/j.mam.2005.12.007

[9]

Molecular diagnosis of α-thalassemia by combining real-time PCR with SYBR Green1 and dissociation curve analysis [J].

Liu, Jingzhong ;

Yan, Mei ;

Wang, Zhangyong ;

Wang, Lirong ;

Zhou, Yan ;

Xiao, Bai .

TRANSLATIONAL RESEARCH, 2006, 148 (01) :6-12

[10]

LIVAK KJ, 1995, PCR METH APPL, V4, P357

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