中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2012年
7期
634-638
,共5页
许笑%张心菊%吴之源%许小平%陈波斌%胡婷婷%陈宇明%关明
許笑%張心菊%吳之源%許小平%陳波斌%鬍婷婷%陳宇明%關明
허소%장심국%오지원%허소평%진파빈%호정정%진우명%관명
血小板增多%骨髓纤维化%受体,血小板生成素%Janus激酶2%突变%聚合酶链反应
血小闆增多%骨髓纖維化%受體,血小闆生成素%Janus激酶2%突變%聚閤酶鏈反應
혈소판증다%골수섬유화%수체,혈소판생성소%Janus격매2%돌변%취합매련반응
Thrombocytosis%Myelofibrosis%Receptors,thrombopoietin%Janus kinase 2%Mutation%Polymerase chain reaction
目的 建立一种能够简便、灵敏地检测原发性血小板增多症(ET)及原发性骨髓纤维化(PMF)患者外周血MPL515突变的方法,探讨我国ET和PMF患者的MPL5I5突变频率及JAK2V617F突变情况.方法 选取2008至2010年复旦大学附属华山医院ET患者261例和PMF患者25例,提取患者外周血DNA.使用SYBR Green Ⅰ荧光定量PCR检测标本的JAK2V617F突变.分别采用针对MPL515wt、MPIW515L和MPLW515K这3种基因型设计的Taqman探针,使用荧光定量PCR法检测MPL515突变,并通过T-A克隆挑取纯克隆测序确认.结果 在261例ET患者中,检测到JAK2V617F突变119例(45.6％),MPL515突变7例(2.7％),包括5例MPLW515L突变、1例MPLW515K突变和1例MPLW515L+K突变.25例PMF患者中,JAK2V617F突变为10例(40.0％),M PL515突变为3例(12.0％),包括1例MPLW515L突变和2例MPLW515L+K突变.1例M PLW515K突变的ET患者同时伴有JAK2V617F突变.结论 JAK2V617F是ET和PMF的分子标志物,而在JAK2V617F阴性的病例中,MPL515突变则是一个重要、有益的补充.
目的 建立一種能夠簡便、靈敏地檢測原髮性血小闆增多癥(ET)及原髮性骨髓纖維化(PMF)患者外週血MPL515突變的方法,探討我國ET和PMF患者的MPL5I5突變頻率及JAK2V617F突變情況.方法 選取2008至2010年複旦大學附屬華山醫院ET患者261例和PMF患者25例,提取患者外週血DNA.使用SYBR Green Ⅰ熒光定量PCR檢測標本的JAK2V617F突變.分彆採用針對MPL515wt、MPIW515L和MPLW515K這3種基因型設計的Taqman探針,使用熒光定量PCR法檢測MPL515突變,併通過T-A剋隆挑取純剋隆測序確認.結果 在261例ET患者中,檢測到JAK2V617F突變119例(45.6％),MPL515突變7例(2.7％),包括5例MPLW515L突變、1例MPLW515K突變和1例MPLW515L+K突變.25例PMF患者中,JAK2V617F突變為10例(40.0％),M PL515突變為3例(12.0％),包括1例MPLW515L突變和2例MPLW515L+K突變.1例M PLW515K突變的ET患者同時伴有JAK2V617F突變.結論 JAK2V617F是ET和PMF的分子標誌物,而在JAK2V617F陰性的病例中,MPL515突變則是一箇重要、有益的補充.
목적 건립일충능구간편、령민지검측원발성혈소판증다증(ET)급원발성골수섬유화(PMF)환자외주혈MPL515돌변적방법,탐토아국ET화PMF환자적MPL5I5돌변빈솔급JAK2V617F돌변정황.방법 선취2008지2010년복단대학부속화산의원ET환자261례화PMF환자25례,제취환자외주혈DNA.사용SYBR Green Ⅰ형광정량PCR검측표본적JAK2V617F돌변.분별채용침대MPL515wt、MPIW515L화MPLW515K저3충기인형설계적Taqman탐침,사용형광정량PCR법검측MPL515돌변,병통과T-A극륭도취순극륭측서학인.결과 재261례ET환자중,검측도JAK2V617F돌변119례(45.6％),MPL515돌변7례(2.7％),포괄5례MPLW515L돌변、1례MPLW515K돌변화1례MPLW515L+K돌변.25례PMF환자중,JAK2V617F돌변위10례(40.0％),M PL515돌변위3례(12.0％),포괄1례MPLW515L돌변화2례MPLW515L+K돌변.1례M PLW515K돌변적ET환자동시반유JAK2V617F돌변.결론 JAK2V617F시ET화PMF적분자표지물,이재JAK2V617F음성적병례중,MPL515돌변칙시일개중요、유익적보충.
Objective To establish a simple and sensitive method to detect MPL515 mutations in peripheral blood of ET and PMF patients,and investigate the frequencies of the MPL515 and JAK2V617F mutations in Chinese patients.Methods Totallv 261 patients of ET and 25 PMF cases were collected from Huashan Hospital of Fudan University and DNA samples were isolated from peripheral blood of these cases.SYBR Green Ⅰ real-time PCR was used to detect JAK2V617F mutation.Taqman probe was designed to be specific for the three types of mutations ( MPl515wt,MPLW515L and MPIW515K).Real-time PCR was used to detect MPL515 mutations.Tbe results were confirmed by sequencing after T-A cloning.Results Among 261 ET patients,119 cases (45.6％ ) were identified as JAK2V617F mutation carriers and 7 cases (2.7％ ) were detected to be MPl515 mutation carriers,including 5 cases with MPLW515L,1 case with MPLW515K and 1 ease with MPLW515L + K.Additionally 10 cases with JAK2V617F(40.0％ ) and 3 cases with MPL515 ( 12.0％ ) were screened out in 25 PMF patients,including 1 case with MPLW515L and 2 cases with MPLW515L + K.One ET patient was found to harbor concurrent JAK2V617F and MPL515 mutations.Conclusion JAK2V617F mutation is the major molecular marker of ET and PMF,meanwhile MPL515 mutation is important and useful complement.
