PCR-SSCP检测肥厚型心肌病心肌肌钙蛋白Ⅰ基因7号外显子突变

Detection of mutation in exon 7 of cardiac troponin Ⅰ gene in patients with hypertrophic cardiomyopathy by PCR-SSCP

目的研究聚合酶链反应-单链构象多态性(PCR-SSCP)法检测肥厚型心肌病心肌肌钙蛋白I(cTnI)基因7号外显子突变的可行性。方法肥厚型心肌病患者59例(2例曾直接测序证实7号外显子Arg145Trp突变),PCR扩增cTnI基因7号外显子,予PCR-SSCP检测突变,与直接测序结果进行比较。结果2例Arg145Trp突变者均有异常条带,余未证实突变者未见阳性条带。结论PCR-SSCP检测肥厚型心肌病cTnI基因Arg145Trp突变具有良好的检出性,可作为大规模人群筛查的工具。

Objective To screen for mutation in exon 7 of cardiac troponin Ⅰ （cTnⅠ） gene in patients with hypertrophic cardiomyopathy by PCR-single strand conformation polymorphism （SSCP） analysis. Methods Fifty-nine patients with hypertrophic cardiomyopathy （cTnⅠ gene Arg145Trp mutation was identified Jn exon 7 in 2 out of 59 patients in previous direct sequencing study） were enrolled and genomic DNA was prepared from their peripheral blood leukocytes. Exons 7 of cTnⅠ gene in patients were amplified by PCR, and products were analyzed by SSCP technique. Results The two patients with Arg145Trp mutation had different SSCP patterns as compared with other patients. There were no false positives found in the rest of patients. Conclusion PCR-SSCP is capable of detecting Arg145Trp mutation in exon 7 of cTnⅠ gene in patients with hypertrophic cardiomyopathy. It may be an useful tool for large-scale screening of this kind of mutation.