摘要
目的探讨糖原累积病Ⅱ型的酶学诊断方法,并在中国人中进行基因突变分析。方法对1例临床表现为婴儿型糖原累积病Ⅱ型的8个月男性患儿进行研究,采用荧光分光光度计检测外周血白细胞酸性α#葡萄糖苷酶(GAA)活性,并采用16对引物对该家系GAA基因从外显子2到外显子20的编码区进行扩增并进行测序分析。结果该患儿白细胞GAA活性仅为18.72nmol/(mgprotein·hr),正常对照组为(130.5±86.7)nmol/(mgprotein·hr);基因学分析发现该患儿存在2个杂合突变位点,其中之一为1935位点C→A错义突变,是中国人最常见的突变位点,另外一个为988位点T→G错义突变,该位点为新发现突变位点。结论糖原累积病Ⅱ型是由于GAA基因突变引起GAA活性降低所致,酶活性检测和基因检测是目前最有效和可靠的诊断方法。
Objectives To investigate the biochemical diagnostic method of glycogen storage disease type Ⅱ （GSD Ⅱ ） and the gene mutations in an Chinese infant with this disease. Methods One 8 month-old boy presented as infantile type of GSD Ⅱ was studied. Acid alpha-D-glucosidase （GAA） activity in the peripheral blood leucocytes was measured using a fluorescence mieroplate reader to confirm the diagnosis. Sixteen pairs of intronie oligonucleotide primer were used to amplify the coding regions of GAA gene from exon 2 to exon 20 in the family members. The coding regions were sequenced both in the sense and antisense strands. Results The leukocytes GAA activity in this infant （18.72 nmol/（mg protein· hr）） was considerably lower than that in normal controls （130.5 ± 86.7 nmol/（mg protein· hr））. Genomic DNA sequencing revealed that the patient was heterozygous for 2 missense mutations. One allele con- rained the C1935A transversion which was the commonest mutation in Chinese patients,with the other containing a novel mutation of the T988G transversion. Conclusions GSD Ⅱ is caused by the deficiency of acid alpha-D-glucosidase,resulting from the mutations in the GAA gene. Measuring the level of enzyme activity and detection of the genomic mutations are the most accurate and definitive diagnostic method available at present.
出处
《临床儿科杂志》
CAS
CSCD
北大核心
2006年第12期962-965,共4页
Journal of Clinical Pediatrics
关键词
Ⅱ型糖原累积病
酸性α-葡萄糖苷酶
基因突变
glycogen storage disease type Ⅱ
acid alpha-D-glucosidase
genomic mutation
