背景:作为血小板上的主要抗原之一,CD36抗原又被称为血小板糖蛋白Ⅳ,其基因变异会导致CD36抗原缺失。目的:序列分析并确认1例CD36抗原新等位基因。方法:提取外周血样本DNA,应用聚合酶链式反应扩增CD36基因的12个编码区序列片段,应用直...背景:作为血小板上的主要抗原之一,CD36抗原又被称为血小板糖蛋白Ⅳ,其基因变异会导致CD36抗原缺失。目的:序列分析并确认1例CD36抗原新等位基因。方法:提取外周血样本DNA,应用聚合酶链式反应扩增CD36基因的12个编码区序列片段,应用直接测序法对目的片段的序列进行检测。所得序列与基因库中编号为NG_008192的标准序列进行比对分析,以确定新的基因突变。结果与结论:被检样本在第12外显子的1142位发生T>G的碱基突变,其他外显子序列与标准序列一致。检索国际基因数据库Gen Bank和美国国立生物技术信息中心(National Center for Biotechnology Information,NCBI),均未发现关于1142 T>G突变的数据和报道,因此为国际上首次确认,上报Gen Bank获得登录号:KM275213。1142 T>G突变导致第381位氨基酸由亮氨酸(Leu,L)改变为丝氨酸(Ser,S),两者在亲/疏水性和极性上差异较大,且381位氨基酸所在区域为高度保守区域,因此推测该突变会使蛋白活性降低甚至消失。展开更多
Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in ...Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in intron 1 and Parkinson’s disease （PD） .Methods We screened all 7exons and exon-intron boundary regions of DJ-1 by PCR and direct nucleotide sequencing in 90 Chinese patients with EOPD.We also compared the allele and genotype frequencies of the g.168<sub>1</sub>85del polymorphism展开更多
Conventional PCR methods combined with linkage analysis based on short tandem repeats （STRs） or Karyomapping with single nucleotide polymorphism （SNP） arrays, have been applied to preimplantation genetic diagnosis...Conventional PCR methods combined with linkage analysis based on short tandem repeats （STRs） or Karyomapping with single nucleotide polymorphism （SNP） arrays, have been applied to preimplantation genetic diagnosis （PGD） for spinal muscular atrophy （SMA）, an autosome recessive disorder. However, it has limitations in SMA diagnosis by Karyomapping, and these methods are unable to distinguish wild- type embryos with carriers effectively. Mutated allele revealed by sequencing with aneuploidy and linkage analyses （MARSALA） is a new method allowing embryo selection by a one-step next-generation sequencing （NGS） procedure, which has been applied in PGD for both autosome dominant and X-linked diseases in our group previously. In this study, we carried out PGD based on MARSALA for two carrier families with SMA affected children. As a result, one of the couples has given birth to a healthy baby free of mutations in SMA-causing gene. It is the first time that MARSALA was applied to PGD for SMA, and we can distinguish the embryos with heterozygous deletion （carriers） from the wild-type （normal） ones accurately through this NGS-based method. In addition, direct mutation detection allows us to identify the affected embryos （homozygous deletion）, which can be regarded as probands for linkage analysis, in case that the affected family member is absent, In the future, the NGS-based MARSALA method is expected to be used in PGD for all monogenetic disorders with known pathogenic gene mutation.展开更多
目的鉴定1例人类白细胞抗原（human leukocyte antigen，HLA）新等位基因。方法应用双链及单链测序法进行基于序列的分型（sequence based typing，SBT），通过群体调查了解该等位基因的群体分布频率。结果该等位基因HLA—A＊33：44，与...目的鉴定1例人类白细胞抗原（human leukocyte antigen，HLA）新等位基因。方法应用双链及单链测序法进行基于序列的分型（sequence based typing，SBT），通过群体调查了解该等位基因的群体分布频率。结果该等位基因HLA—A＊33：44，与A＊33：03：01第4外显子相差1个核苷酸，使第866位碱基由G变为A，第265个密码子由GGT变为GAT，相应氨基酸由甘氨酸变为天门冬氨酸。与IMGT／HLA数据库中的HLA—A等位基因的序列进行对比，该突变为新的单核苷酸多态性位点。结论该新HLA等位基因在中国汉族人群中的分布频率小于0．0003，并被世界卫生组织HLA因子命名委员会命名为HLA—A＊33：44（序列注册号为HQ873871）。研究新等位基因的序列可为HLA基因相关研究和应用提供信息。展开更多
A silent mutation or sequence polymorphism, A to T substitution at codon 399 in exon11 of the PAH gene from a Chinese PKU patient, was found by sequence analysis. The fre-quencies of this new mutation in normal and ab...A silent mutation or sequence polymorphism, A to T substitution at codon 399 in exon11 of the PAH gene from a Chinese PKU patient, was found by sequence analysis. The fre-quencies of this new mutation in normal and abnormal (PKU) genes were 0.005 and 0.09,respectively, based on the analyses of 100 normal individuals and 39 PKU patients usingDNA amplification with polymerase chain reaction (PCR) and oligonucleotide hybridizationmethods. This silent mutation can be used as a "genetic marker" for PKU prenatal diagno-sis. Recently, a fetus at risk for PKU, who could not be completely predicted by RFLPslinkage analysis, was prenatally diagnosed with this genetic marker.展开更多
Objective To describe the clinical characteristics,and to analyze the AGXT gene mutation in three siblings with primary hyperoxaluria typeⅠ(PHI).Methods AGXT gene mutation was analyzed by direct sequencing analysis i...Objective To describe the clinical characteristics,and to analyze the AGXT gene mutation in three siblings with primary hyperoxaluria typeⅠ(PHI).Methods AGXT gene mutation was analyzed by direct sequencing analysis in this family,and the minor allele status was also tested.One hundred unrelated healthy subjects were also analyzed as controls.Results Three mutations展开更多
Objective To explore the association between LMNA gene mutation and familiar dilated cardiomyopathy(DCM)(FDCM)and idiopathic DCM(IDCM)in Uygurs and Han people in Xinjiang area.Methods Peripheral blood samples were col...Objective To explore the association between LMNA gene mutation and familiar dilated cardiomyopathy(DCM)(FDCM)and idiopathic DCM(IDCM)in Uygurs and Han people in Xinjiang area.Methods Peripheral blood samples were collected from 28 family member with FDCM and 123 sporadic patients with IDCM(56 Uygur patients and 67 Han patients),80展开更多
文摘背景:作为血小板上的主要抗原之一,CD36抗原又被称为血小板糖蛋白Ⅳ,其基因变异会导致CD36抗原缺失。目的:序列分析并确认1例CD36抗原新等位基因。方法:提取外周血样本DNA,应用聚合酶链式反应扩增CD36基因的12个编码区序列片段,应用直接测序法对目的片段的序列进行检测。所得序列与基因库中编号为NG_008192的标准序列进行比对分析,以确定新的基因突变。结果与结论:被检样本在第12外显子的1142位发生T>G的碱基突变,其他外显子序列与标准序列一致。检索国际基因数据库Gen Bank和美国国立生物技术信息中心(National Center for Biotechnology Information,NCBI),均未发现关于1142 T>G突变的数据和报道,因此为国际上首次确认,上报Gen Bank获得登录号:KM275213。1142 T>G突变导致第381位氨基酸由亮氨酸(Leu,L)改变为丝氨酸(Ser,S),两者在亲/疏水性和极性上差异较大,且381位氨基酸所在区域为高度保守区域,因此推测该突变会使蛋白活性降低甚至消失。
文摘Objective To evaluate the prevalence of the DJ-1mutation in early-onset Parkinson’s disease （EOPD） patients,and analyzed the association between the certain polymorphic marker g.168<sub>1</sub>85del in intron 1 and Parkinson’s disease （PD） .Methods We screened all 7exons and exon-intron boundary regions of DJ-1 by PCR and direct nucleotide sequencing in 90 Chinese patients with EOPD.We also compared the allele and genotype frequencies of the g.168<sub>1</sub>85del polymorphism
基金supported by the National Natural Science Foundation of China (Nos. 31522034, 31571544 and 31230047)the National High Technology Research and Development Program (No. 2015AA020407)+1 种基金Beijing Municipal Science and Technology Commission (No. D151100002415004)Research Fund of National Health and Family Planning Commission of China (No. 201402004)
文摘Conventional PCR methods combined with linkage analysis based on short tandem repeats （STRs） or Karyomapping with single nucleotide polymorphism （SNP） arrays, have been applied to preimplantation genetic diagnosis （PGD） for spinal muscular atrophy （SMA）, an autosome recessive disorder. However, it has limitations in SMA diagnosis by Karyomapping, and these methods are unable to distinguish wild- type embryos with carriers effectively. Mutated allele revealed by sequencing with aneuploidy and linkage analyses （MARSALA） is a new method allowing embryo selection by a one-step next-generation sequencing （NGS） procedure, which has been applied in PGD for both autosome dominant and X-linked diseases in our group previously. In this study, we carried out PGD based on MARSALA for two carrier families with SMA affected children. As a result, one of the couples has given birth to a healthy baby free of mutations in SMA-causing gene. It is the first time that MARSALA was applied to PGD for SMA, and we can distinguish the embryos with heterozygous deletion （carriers） from the wild-type （normal） ones accurately through this NGS-based method. In addition, direct mutation detection allows us to identify the affected embryos （homozygous deletion）, which can be regarded as probands for linkage analysis, in case that the affected family member is absent, In the future, the NGS-based MARSALA method is expected to be used in PGD for all monogenetic disorders with known pathogenic gene mutation.
文摘目的鉴定1例人类白细胞抗原（human leukocyte antigen，HLA）新等位基因。方法应用双链及单链测序法进行基于序列的分型（sequence based typing，SBT），通过群体调查了解该等位基因的群体分布频率。结果该等位基因HLA—A＊33：44，与A＊33：03：01第4外显子相差1个核苷酸，使第866位碱基由G变为A，第265个密码子由GGT变为GAT，相应氨基酸由甘氨酸变为天门冬氨酸。与IMGT／HLA数据库中的HLA—A等位基因的序列进行对比，该突变为新的单核苷酸多态性位点。结论该新HLA等位基因在中国汉族人群中的分布频率小于0．0003，并被世界卫生组织HLA因子命名委员会命名为HLA—A＊33：44（序列注册号为HQ873871）。研究新等位基因的序列可为HLA基因相关研究和应用提供信息。
文摘A silent mutation or sequence polymorphism, A to T substitution at codon 399 in exon11 of the PAH gene from a Chinese PKU patient, was found by sequence analysis. The fre-quencies of this new mutation in normal and abnormal (PKU) genes were 0.005 and 0.09,respectively, based on the analyses of 100 normal individuals and 39 PKU patients usingDNA amplification with polymerase chain reaction (PCR) and oligonucleotide hybridizationmethods. This silent mutation can be used as a "genetic marker" for PKU prenatal diagno-sis. Recently, a fetus at risk for PKU, who could not be completely predicted by RFLPslinkage analysis, was prenatally diagnosed with this genetic marker.
文摘Objective To describe the clinical characteristics,and to analyze the AGXT gene mutation in three siblings with primary hyperoxaluria typeⅠ(PHI).Methods AGXT gene mutation was analyzed by direct sequencing analysis in this family,and the minor allele status was also tested.One hundred unrelated healthy subjects were also analyzed as controls.Results Three mutations
文摘Objective To explore the association between LMNA gene mutation and familiar dilated cardiomyopathy(DCM)(FDCM)and idiopathic DCM(IDCM)in Uygurs and Han people in Xinjiang area.Methods Peripheral blood samples were collected from 28 family member with FDCM and 123 sporadic patients with IDCM(56 Uygur patients and 67 Han patients),80
