Gu Feng, Shi Yifan, Deng Jieying, Jin Zimeng
Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing 100730, China.
Zhonghua Yi Xue Za Zhi. 2002 Oct 25;82(20):1401-5.
To investigate the mutations of arginine vasopressin receptor 2 (AVPR2) gene in congenital nephrogenic diabetes insipidus (NDI) patients.
Genomic DNA was extracted from the blood samples of 7 congenital NDI patients, all males, and 24 of their family members. The 6 fragments of AVPR2 gene were amplified by PCR. The gene mutations were preliminarily screened by SSCP and further confirmed by DNA sequencing.
5 types of mutation and 8 mutant sites of AVPR2 gene were found in 6 of the 7 patients. Two cases were found to have 2 different sites of mutation in the AVPR2 gene. In 4 cases new mutation sites not reported previously in the literature, i.e., g.469-493del 24, g.541insT, g.462delC and g.935T > C, were identified, resulting in the change of AVPR2 proteins, including A37-L44del (deletion mutation), A61G 190X (insertion frame-shift and nonsense mutation), P34R 36X (deletion frame-shift and nonsense mutation) and C192R (missense mutation). Among the 24 family members examined only one, a mother, was found to have an AVPR2 gene mutation with the same mutation site as the proband.
Four novel mutation sites have been identified in the AVPR2 gene among the congenital NDI patients. PCR-SSCP and DNA sequencing can be used to preliminarily screen and diagnose gene mutation among congenital NDI patients.
研究先天性肾性尿崩症(NDI)患者精氨酸加压素受体2(AVPR2)基因的突变情况。
从7例先天性NDI患者(均为男性)及其24名家庭成员的血液样本中提取基因组DNA。通过聚合酶链反应(PCR)扩增AVPR2基因的6个片段。采用单链构象多态性(SSCP)初步筛选基因突变,并用DNA测序进一步确认。
7例患者中的6例发现了5种AVPR2基因突变类型和8个突变位点。2例患者在AVPR2基因中发现了2个不同的突变位点。4例患者中发现了文献中未曾报道的新突变位点,即g.469 - 493del 24、g.541insT、g.462delC和g.935T > C,这些突变导致了AVPR2蛋白的改变,包括A37 - L44del(缺失突变)、A61G 190X(插入移码和无义突变)、P34R 36X(缺失移码和无义突变)和C192R(错义突变)。在检测的24名家庭成员中,仅1名母亲被发现与先证者具有相同的AVPR2基因突变位点。
在先天性NDI患者中,AVPR2基因鉴定出4个新的突变位点。PCR - SSCP和DNA测序可用于先天性NDI患者基因突变的初步筛选和诊断。