Most common SLC25A13 mutation in 400 Chinese infants with intrahepatic cholestasis
Most common SLC25A13 mutation in 400 Chinese infants with intrahepatic cholestasis
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400名中国婴儿肝内胆汁淤积症最常见的SLC25A13突变
DOI：
10.3748/wjg.v16.i18.2278
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发表时间：
2010-05-14
影响因子：
4.3
通讯作者：
Wang, Jian-She
中科院分区：
文献类型：
--
作者：
Fu, Hai-Yan;Zhang, Shao-Ren;Wang, Jian-She
目的：建立双标记探针实时荧光聚合酶链反应（RT-PCR）快速检测SLC25A13基因突变851del4的方法。方法：选取来自中国18个省市的400例不明原因肝内胆汁淤积症婴儿（小于1岁），检测其SLC25A13基因突变851del4。设计了适合SLC25A13基因突变841del4的引物和荧光标记探针。用两个荧光标记探针进行正常和突变序列的PCR检测。单次RT-PCR后，通过分析起飞曲线得到结果。24份阳性样本和14份阴性样本采用直接测序法重新检测。结果：在46个突变等位基因中检测到8个纯合突变和30个杂合突变，851del4突变率为5.8%（46/800）。在中国中部和南部地区的474个和242个等位基因中，分别观察到26个和20个突变等位基因。中国北方84个等位基因中未检测到突变等位基因。对24份阳性样本（纯合突变4份，杂合突变20份）和14份阴性样本进行直接测序重测，证实RTPCR的准确性为100%。结论：RT-PCR检测婴儿肝内胆汁淤积症中851del4基因突变的准确率为100%。(C) 2010年白石登。版权所有。
AIM: To establish the real time fluorescence polymerase chain reaction (RT-PCR) with dual labeled probes for fast detection of SLC25A13 gene mutation 851del4.METHODS: Four hundred infants (< 1 year of age) with unexplained intrahepatic cholestasis from 18 provinces or municipalities in China were enrolled in this study for detecting their SLC25A13 gene mutation 851del4. Suitable primers and fluorescence-labeled probes for detecting SLC25A13 gene mutation 841del4 were designed. Normal and mutant sequences were detected by PCR with two fluorescence-labeled probes. After a single RT-PCR, results were obtained by analyzing the take-off curves. Twenty-four positive and 14 negative samples were retested by direct sequencing.RESULTS: Eight homozygous and 30 heterozygous mutations were detected in 46 mutant alleles with a 851del4 mutation rate of 5.8% (46/800). Twenty-six and 20 mutant alleles were observed respectively, in 474 and 242 alleles from the intermediate and southern areas of China. No mutant allele was detected in 84 alleles from northern China. Twenty-four positive samples including 4 homozygous and 20 heterozygous mutations, and 14 negative samples were retested by direct sequencing, which confirmed that the accuracy of RTPCR was 100%.CONCLUSION: RT-PCR can detect the mutation 851del4 in infants with intrahepatic cholestasis with an accuracy of 100%. (C) 2010 Baishideng. All rights reserved.