Zheng Bixia, Wang Chunli, Jia Zhanjun, Liu Zhifeng, Li Mei, Jin Yu, Pan Jian
*Nanjing Key Laboratory of Pediatrics †Department of Gastroenterology, Nanjing Children's Hospital, Nanjing Medical University, Nanjing 210008, China.
J Pediatr Gastroenterol Nutr. 2017 Apr;64(4):559-564. doi: 10.1097/MPG.0000000000001316.
Peutz-Jeghers syndrome (PJS) is caused by the germline mutations in serine/threonine kinase 11 (STK11) gene. The aim of the present study was to investigate the spectrum of STK11 gene mutations using multiplex ligation-dependent probe amplification (MLPA) assay in combination with direct sequencing in Chinese children with PJS.
Nine children who met the clinical criteria for PJS and 2 presumed patients with PJS were enrolled in the present study. Patients' clinical information on polyp characteristics, polyp-related complications, family histories, and so on were reviewed and analyzed. After obtaining informed consent, we performed a mutation analysis of STK11 gene in 11 Chinese patients using MLPA assay and direct sequencing.
By means of MLPA method, we detected exonic deletions in 5 patients. In details, 1 patient had the complete deletion of all 10 exons, 3 patients showed deletions of promoter region and exon 1, and 1 patient had exon deletions from 1 to 9. By direct sequencing of the coding region of STK11 gene, we identified point mutations in 4 patients at c.548T>G/p.Leu183Arg, c.580G>T/p.Asp194Tyr, c.152_153insGG/Asp53GlyfsX12, and c.631delC/Arg211GlyfsX76, respectively, and 3 of them are novel mutations. We failed to find any mutation in left 2 patients who met the clinical criteria of PJS.
MLPA plus direct sequencing revealed large genomic deletions of STK11 gene in Chinese children with PJS and increased the detecting rate of STK11 gene mutations in Chinese patients with PJS. MLPA combined with direct sequencing could serve as a better strategy for the genetic diagnosis of PJS in Chinese population.
黑斑息肉综合征（PJS）由丝氨酸/苏氨酸激酶11（STK11）基因的种系突变引起。本研究旨在采用多重连接依赖探针扩增（MLPA）分析结合直接测序法，研究中国PJS患儿中STK11基因突变谱。
本研究纳入9例符合PJS临床标准的患儿以及2例疑似PJS患者。回顾并分析了患者关于息肉特征、息肉相关并发症、家族史等的临床信息。在获得知情同意后，我们采用MLPA分析和直接测序法对11例中国患者的STK11基因进行了突变分析。
通过MLPA方法，我们在5例患者中检测到外显子缺失。具体而言，1例患者所有10个外显子均完全缺失，3例患者显示启动子区域和外显子1缺失，1例患者外显子1至9缺失。通过对STK11基因编码区的直接测序，我们分别在4例患者中鉴定出c.548T>G/p.Leu183Arg、c.580G>T/p.Asp194Tyr、c.152_153insGG/Asp53GlyfsX12和c.631delC/Arg211GlyfsX76的点突变，其中3个为新突变。我们在另外2例符合PJS临床标准的患者中未发现任何突变。
MLPA结合直接测序揭示了中国PJS患儿中STK11基因存在大片段基因组缺失，并提高了中国PJS患者中STK11基因突变的检测率。MLPA联合直接测序可作为中国人群PJS基因诊断的更好策略。
