国际输血及血液学杂志
國際輸血及血液學雜誌
국제수혈급혈액학잡지
INTERNATIONAL JOURNAL OF BLOOD TRANSFUSION AND HEMATOLOGY
2013年
2期
102-105
,共4页
GPA%GPB%GYPA%GYPB%基因测序%基因遗传
GPA%GPB%GYPA%GYPB%基因測序%基因遺傳
GPA%GPB%GYPA%GYPB%기인측서%기인유전
GPA%GPB%GYPA%GYPB%gene sequencing%genetic
目的 建立人类GPA,GPB分子相关基因GYPA,GYPB外显子全长序列的DNA直接测序方法,并将此方法应用于RBC血型MNS抗原的分析.方法 以整群随机抽样法选择从2011年11月至2012年5月在本中心无偿献血的175例健康成年人的全血样品为研究对象.以血清学方法与序列特异引物聚合酶链反应(PCR-SSP)方法鉴定其MNS血型.根据GenBank提供的NG-007470与NG-007483基因参照序列为依据,自行设计GPA相关基因GYPA全部外显子的7对引物与GPB相关基因GYPB全部外显子的5对引物,探索最佳反应体系与PCR扩增条件,且能同时扩增2个基因全长外显子碱基序列,使用ABI3730XL DNA测序仪进行序列分析,以Oli6installer分析软件分析碱基序列.结果 所建立的测序方法可于同一反应体系、相同PCR扩增条件同时检测GPA,GPB相关基因GYPA,GYPB外显子全长序列,并准确分析本组血液样本的MNS血型的特征.基因测序结果与血清学分析及PCR-SSP鉴定结果完全一致.结论 首次建立了稳定、精确、简捷的基因直接测序分型方法,可用于检测MNS血型基因型,并分析GPA,GPB肽链中氨基酸序列的多态性,为MNS血型系统的研究以及人类群体遗传学及分子进化的研究等方面提供广泛的应用价值.
目的 建立人類GPA,GPB分子相關基因GYPA,GYPB外顯子全長序列的DNA直接測序方法,併將此方法應用于RBC血型MNS抗原的分析.方法 以整群隨機抽樣法選擇從2011年11月至2012年5月在本中心無償獻血的175例健康成年人的全血樣品為研究對象.以血清學方法與序列特異引物聚閤酶鏈反應(PCR-SSP)方法鑒定其MNS血型.根據GenBank提供的NG-007470與NG-007483基因參照序列為依據,自行設計GPA相關基因GYPA全部外顯子的7對引物與GPB相關基因GYPB全部外顯子的5對引物,探索最佳反應體繫與PCR擴增條件,且能同時擴增2箇基因全長外顯子堿基序列,使用ABI3730XL DNA測序儀進行序列分析,以Oli6installer分析軟件分析堿基序列.結果 所建立的測序方法可于同一反應體繫、相同PCR擴增條件同時檢測GPA,GPB相關基因GYPA,GYPB外顯子全長序列,併準確分析本組血液樣本的MNS血型的特徵.基因測序結果與血清學分析及PCR-SSP鑒定結果完全一緻.結論 首次建立瞭穩定、精確、簡捷的基因直接測序分型方法,可用于檢測MNS血型基因型,併分析GPA,GPB肽鏈中氨基痠序列的多態性,為MNS血型繫統的研究以及人類群體遺傳學及分子進化的研究等方麵提供廣汎的應用價值.
목적 건립인류GPA,GPB분자상관기인GYPA,GYPB외현자전장서렬적DNA직접측서방법,병장차방법응용우RBC혈형MNS항원적분석.방법 이정군수궤추양법선택종2011년11월지2012년5월재본중심무상헌혈적175례건강성년인적전혈양품위연구대상.이혈청학방법여서렬특이인물취합매련반응(PCR-SSP)방법감정기MNS혈형.근거GenBank제공적NG-007470여NG-007483기인삼조서렬위의거,자행설계GPA상관기인GYPA전부외현자적7대인물여GPB상관기인GYPB전부외현자적5대인물,탐색최가반응체계여PCR확증조건,차능동시확증2개기인전장외현자감기서렬,사용ABI3730XL DNA측서의진행서렬분석,이Oli6installer분석연건분석감기서렬.결과 소건립적측서방법가우동일반응체계、상동PCR확증조건동시검측GPA,GPB상관기인GYPA,GYPB외현자전장서렬,병준학분석본조혈액양본적MNS혈형적특정.기인측서결과여혈청학분석급PCR-SSP감정결과완전일치.결론 수차건립료은정、정학、간첩적기인직접측서분형방법,가용우검측MNS혈형기인형,병분석GPA,GPB태련중안기산서렬적다태성,위MNS혈형계통적연구이급인류군체유전학급분자진화적연구등방면제공엄범적응용개치.
Objective To establish ot human GPA,GPB molecules related gene GYPA,GYPB exon full-length sequences of DNA direct sequencing method and applied to red cell blood group MNS antigen analysis.Methods From November 2011 to May 2012,175 whole blood samples from healthy adult donors were included in this study by cluster random sampling method.MNS blood types were identified by serological method and the PCR-SSP method,respectively.According to the GenBank NG-007470 and NG-007483 gene sequence as the basis,the GPA related gene GYPA all exons of the 7 pairs of primers and GPB related gene GYPB all exons of 5 pairs of primers were designed.We had explored the optimum reaction system with the PCR conditions that could amplify the full-length genes on the simultaneous.The base sequence was checked by the PrismTMABI3730XL DNA sequencer and the Oli6installer analysis software.Results The present study established a stable,accurate,simple method that could direct all exons length sequence of the GPA,GPB related genes GYPA,GYPB and propagate all the DNA sequence in the same reaction system,the same PCR amplification conditions for simultaneous.All of the 175 samples of MNS blood group characteristics were accurate analysis by the gene sequencing method.The results of DNA sequencing,serological analysis and PCR-SSP identification results were completely consistent.Conclusions This method was established in the same conditions for detection all exons of the gene full-length sequences about the GPA,GPB molecules related gene GYPA,GYPB for the first time.The method could detect the MNS blood group genotype and analysis of GPA,GPB molecular of amino acid sequence polymorphism.The method has the extensive application value about the MNS blood group system research as well as the human population genetics and molecular evolution research areas.
