Molecular diagnostic testing for Klinefelter syndrome and other male sex chromosome aneuploidies.
Molecular diagnostic testing for Klinefelter syndrome and other male sex chromosome aneuploidies.
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DOI：
10.1186/1687-9856-2012-8
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发表时间：
2012-04-23
期刊：
影响因子：
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通讯作者：
Rinder HM
中科院分区：
文献类型：
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作者：
Hager K;Jennings K;Hosono S;Howell S;Gruen JR;Rivkees SA;Tartaglia NR;Rinder HM
男性性染色体非整倍性诊断不足，尽管伴随的身体和行为表现。目的建立一种无创、快速、高通量的检测男性性染色体非整倍体（包括47，XXY（Klinefelter）、47，XYY、48，XXYY和48，XXXY综合征）的分子诊断方法。该测定法利用三种XYM和四种XA标记物分别询问Y：X和X：常染色体比率。使用从具有非整倍体（n = 117）和46，XY（n = 206）核型的323名男性的群组中分离的基因组DNA对七个标记进行PCR扩增。将所得PCR产物进行焦磷酸测序，一种定量DNA测序方法。受试者工作特征曲线（ROC）被用来建立从正常样本中区分非整倍体的阈值。XYM标记物允许鉴定47，XXY，48，XXXY和47，XYY综合征，在纯化的DNA和口腔拭子样品中具有100%的灵敏度和特异性。48，XXYY核型由来自46，XY的XA标记数据描绘; 43%的X等位基因阈值也允许以100%的灵敏度和特异性检测48，XXYY。X染色体特异性双等位基因SNPs分析表明，45例48，XXYY核型的个体中有43例（96%）具有两条不同的X染色体，而2例（4%）具有重复的X，这提供了48，XXYY可能是由46，XY胚胎早期有丝分裂期间的不分离引起的证据。定量焦磷酸测序技术具有高通量的潜力，可以100%灵敏地检测男性性染色体非整倍体。
Male sex chromosome aneuploidies are underdiagnosed despite concomitant physical and behavioral manifestations. To develop a non-invasive, rapid and high-throughput molecular diagnostic assay for detection of male sex chromosome aneuploidies, including 47,XXY (Klinefelter), 47,XYY, 48,XXYY and 48,XXXY syndromes. The assay utilizes three XYM and four XA markers to interrogate Y:X and X:autosome ratios, respectively. The seven markers were PCR amplified using genomic DNA isolated from a cohort of 323 males with aneuploid (n = 117) and 46,XY (n = 206) karyotypes. The resulting PCR products were subjected to Pyrosequencing, a quantitative DNA sequencing method. Receiver operator characteristic (ROC) curves were used to establish thresholds for the discrimination of aneuploid from normal samples. The XYM markers permitted the identification of 47,XXY, 48,XXXY and 47,XYY syndromes with 100% sensitivity and specificity in both purified DNA and buccal swab samples. The 48,XXYY karyotype was delineated by XA marker data from 46,XY; an X allele threshold of 43% also permitted detection of 48,XXYY with 100% sensitivity and specificity. Analysis of X chromosome-specific biallelic SNPs demonstrated that 43 of 45 individuals (96%) with 48,XXYY karyotype had two distinct X chromosomes, while 2 (4%) had a duplicate X, providing evidence that 48,XXYY may result from nondisjunction during early mitotic divisions of a 46,XY embryo. Quantitative Pyrosequencing, with high-throughput potential, can detect male sex chromosome aneuploidies with 100% sensitivity.