中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2012年
2期
170-173
,共4页
莫宗平%张玲%胡朝晖%冯文莉
莫宗平%張玲%鬍朝暉%馮文莉
막종평%장령%호조휘%풍문리
血红蛋白类,异常%胎儿血红蛋白%血红蛋白病%基因型%核酸扩增技术%聚合酶链反应
血紅蛋白類,異常%胎兒血紅蛋白%血紅蛋白病%基因型%覈痠擴增技術%聚閤酶鏈反應
혈홍단백류,이상%태인혈홍단백%혈홍단백병%기인형%핵산확증기술%취합매련반응
Hemoglobins,abnormal%Fetal hemoglobin%Hemoglobinopathies%Genotype%Nucleic acid amplification techniques%Polymerase chain reaction
目的 对1例异常血红蛋白合并遗传性持续性胎儿血红蛋白增多症(HPFH)患者进行基因分析.方法 患者男,26岁,2010年9月到广州金域医学检验中心进行体检,经血液学分析、血红蛋白电泳、红细胞渗透脆性分析后,疑为异常血红蛋白合并HPFH病患者.应用常规血液学检查、碱性琼脂糖凝胶血红蛋白电泳对先证者进行表型分析；通过对HBB基因直接测序检测异常血红蛋白的突变类型；应用多重连接酶依赖探针扩增技术(MLPA)检测整个β-珠蛋白基因簇有无缺失及缺失的大致范围；应用跨越断裂点聚合酶链反应(gap-PCR)进行跨越断裂点PCR扩增,对此扩增产物进行基因测序分析,并将其结果与参考序列NC_000011.9进行比对,判断具体的断裂位点.结果 通过对HBB基因进行直接测序,发现该异常血红蛋白为Ta-Li( HBB:c.250G＞T),通过MLPA结合gap-PCR以及对产物进行基因测序,发现患者存在β-珠蛋白基因的大片段缺失,缺失范围为NC_000011.9:g.5222878_5250288del,即患者的基因型为异常血红蛋白Ta-Li合并东南亚型遗传性持续性胎儿血红蛋白增多症(SEA-HPFH).结论 联合应用MLPA、gap-PCR和基因测序技术可能有助于明确先证者的基因型.
目的 對1例異常血紅蛋白閤併遺傳性持續性胎兒血紅蛋白增多癥(HPFH)患者進行基因分析.方法 患者男,26歲,2010年9月到廣州金域醫學檢驗中心進行體檢,經血液學分析、血紅蛋白電泳、紅細胞滲透脆性分析後,疑為異常血紅蛋白閤併HPFH病患者.應用常規血液學檢查、堿性瓊脂糖凝膠血紅蛋白電泳對先證者進行錶型分析；通過對HBB基因直接測序檢測異常血紅蛋白的突變類型；應用多重連接酶依賴探針擴增技術(MLPA)檢測整箇β-珠蛋白基因簇有無缺失及缺失的大緻範圍；應用跨越斷裂點聚閤酶鏈反應(gap-PCR)進行跨越斷裂點PCR擴增,對此擴增產物進行基因測序分析,併將其結果與參攷序列NC_000011.9進行比對,判斷具體的斷裂位點.結果 通過對HBB基因進行直接測序,髮現該異常血紅蛋白為Ta-Li( HBB:c.250G＞T),通過MLPA結閤gap-PCR以及對產物進行基因測序,髮現患者存在β-珠蛋白基因的大片段缺失,缺失範圍為NC_000011.9:g.5222878_5250288del,即患者的基因型為異常血紅蛋白Ta-Li閤併東南亞型遺傳性持續性胎兒血紅蛋白增多癥(SEA-HPFH).結論 聯閤應用MLPA、gap-PCR和基因測序技術可能有助于明確先證者的基因型.
목적 대1례이상혈홍단백합병유전성지속성태인혈홍단백증다증(HPFH)환자진행기인분석.방법 환자남,26세,2010년9월도엄주금역의학검험중심진행체검,경혈액학분석、혈홍단백전영、홍세포삼투취성분석후,의위이상혈홍단백합병HPFH병환자.응용상규혈액학검사、감성경지당응효혈홍단백전영대선증자진행표형분석；통과대HBB기인직접측서검측이상혈홍단백적돌변류형；응용다중련접매의뢰탐침확증기술(MLPA)검측정개β-주단백기인족유무결실급결실적대치범위；응용과월단렬점취합매련반응(gap-PCR)진행과월단렬점PCR확증,대차확증산물진행기인측서분석,병장기결과여삼고서렬NC_000011.9진행비대,판단구체적단렬위점.결과 통과대HBB기인진행직접측서,발현해이상혈홍단백위Ta-Li( HBB:c.250G＞T),통과MLPA결합gap-PCR이급대산물진행기인측서,발현환자존재β-주단백기인적대편단결실,결실범위위NC_000011.9:g.5222878_5250288del,즉환자적기인형위이상혈홍단백Ta-Li합병동남아형유전성지속성태인혈홍단백증다증(SEA-HPFH).결론 연합응용MLPA、gap-PCR화기인측서기술가능유조우명학선증자적기인형.
Objective To investigate the genotype of a case with abnormal hemoglobin combined with hereditary persistence of fetal hemoglobin (HPFH).Methods Male patient,26 years old,were suspected abnormal hemoglobin combined with HPFH after receiving medical examination including hematology exmination,hemoglobin electrophoresis,erythrocyte osmotic fragility analysis in Guangzhou Kingmed Diagnostics in September 2010.Routine examination of anemia and hemoglobin electrophoresis at alkaine pH on agarose gels were applied to analyze the phenotype.Direct sequencing of the complete HBB gene was utilized to identify the hemoglobin variant.Multiplex ligation-dependent probe amplification (MLPA) assay was used to identify the presence of β-globin gene cluster deletion.Gap polymerase chain reaction (gap-PCR) was used to amplify the HBB gene fragment across the breakpoint,and the deletion breakpoint was characterized by direct sequencing the gap-PCR product and comparing the sequencing result with the reference sequence NC_000011.9.Results By direct sequencing of the complete HBB gene,the patient in this study was found to carry a hemoglobin Ta-Li (HBB:c.250G ＞ T) mutation.By combining use of MLPA and gap-PCR with gene sequencing,we found that it had a gross deletion in the β-globin gene cluster,the deletion region was NC_000011.9:g.5222878_5250288del.Therefore,the genotype of this subject was SEA-HPFH combined with abnormal hemoglobin Ta-li.Conclusion Combining application of MLPA and gap-PCR with gene sequencing can help to make sure the genotype.
