中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
12期
1119-1124
,共6页
马芳%贾小芳%陈军%姚亚敏%刘晓茜%卢洪洲%张丽军
馬芳%賈小芳%陳軍%姚亞敏%劉曉茜%盧洪洲%張麗軍
마방%가소방%진군%요아민%류효천%로홍주%장려군
抗逆转录病毒治疗,高效%获得性免疫缺陷综合征%蛋白质组%病毒载量%生物学标记
抗逆轉錄病毒治療,高效%穫得性免疫缺陷綜閤徵%蛋白質組%病毒載量%生物學標記
항역전록병독치료,고효%획득성면역결함종합정%단백질조%병독재량%생물학표기
Antiretroviral therapy,highly active%Acquired immunodeficieucy syndrome%Proteome%Viral load%Biological markers
目的 筛选AIDS患者血浆中与HAART疗效相关的特异性生物标志物,为临床疗效评估提供新型检测分子,指导AIDS治疗的预后判断.方法 收集2008年6月至2009年2月由上海市公共卫生临床中心感染一科收集的艾滋病患者血浆,包括病毒学治疗有效组10例(病毒载量＜ 50拷贝/ml)和病毒学治疗失败组11例(病毒载量＞50拷贝/ml),入组患者年龄22 ～ 63岁.血浆样品用Bio-rad公司血浆高丰度蛋白质去除试剂盒处理,去除血浆中的白蛋白和免疫球蛋白IgG.处理后的血浆蛋白质样品通过二维凝胶电泳(2DE)进行分离,采用ImageMaster软件分析获得的电泳图谱,找到与疗效相关的差异蛋白质.差异蛋白质点经过胰酶酶切后,通过在线纳升级反向液相色谱串联电喷雾离子阱质谱分析鉴定.结果 血浆经过高丰度蛋白去除试剂盒处理后,低丰度的蛋白质得到了有效的富集.共发现了6个在治疗有效组和治疗无效组血浆样品中表达差异的蛋白质点.这些差异蛋白质经液相色谱串联质谱分析得到准确鉴定,包括血清转铁蛋白、血清β纤维蛋白原等.结论 通过蛋白质组学研究发现,血清转铁蛋白等蛋白质在艾滋病患者血浆中表达水平与治疗过程中病毒载量相关,它们可能是评价抗病毒治疗疗效的潜在生物标志物.
目的 篩選AIDS患者血漿中與HAART療效相關的特異性生物標誌物,為臨床療效評估提供新型檢測分子,指導AIDS治療的預後判斷.方法 收集2008年6月至2009年2月由上海市公共衛生臨床中心感染一科收集的艾滋病患者血漿,包括病毒學治療有效組10例(病毒載量＜ 50拷貝/ml)和病毒學治療失敗組11例(病毒載量＞50拷貝/ml),入組患者年齡22 ～ 63歲.血漿樣品用Bio-rad公司血漿高豐度蛋白質去除試劑盒處理,去除血漿中的白蛋白和免疫毬蛋白IgG.處理後的血漿蛋白質樣品通過二維凝膠電泳(2DE)進行分離,採用ImageMaster軟件分析穫得的電泳圖譜,找到與療效相關的差異蛋白質.差異蛋白質點經過胰酶酶切後,通過在線納升級反嚮液相色譜串聯電噴霧離子阱質譜分析鑒定.結果 血漿經過高豐度蛋白去除試劑盒處理後,低豐度的蛋白質得到瞭有效的富集.共髮現瞭6箇在治療有效組和治療無效組血漿樣品中錶達差異的蛋白質點.這些差異蛋白質經液相色譜串聯質譜分析得到準確鑒定,包括血清轉鐵蛋白、血清β纖維蛋白原等.結論 通過蛋白質組學研究髮現,血清轉鐵蛋白等蛋白質在艾滋病患者血漿中錶達水平與治療過程中病毒載量相關,它們可能是評價抗病毒治療療效的潛在生物標誌物.
목적 사선AIDS환자혈장중여HAART료효상관적특이성생물표지물,위림상료효평고제공신형검측분자,지도AIDS치료적예후판단.방법 수집2008년6월지2009년2월유상해시공공위생림상중심감염일과수집적애자병환자혈장,포괄병독학치료유효조10례(병독재량＜ 50고패/ml)화병독학치료실패조11례(병독재량＞50고패/ml),입조환자년령22 ～ 63세.혈장양품용Bio-rad공사혈장고봉도단백질거제시제합처리,거제혈장중적백단백화면역구단백IgG.처리후적혈장단백질양품통과이유응효전영(2DE)진행분리,채용ImageMaster연건분석획득적전영도보,조도여료효상관적차이단백질.차이단백질점경과이매매절후,통과재선납승급반향액상색보천련전분무리자정질보분석감정.결과 혈장경과고봉도단백거제시제합처리후,저봉도적단백질득도료유효적부집.공발현료6개재치료유효조화치료무효조혈장양품중표체차이적단백질점.저사차이단백질경액상색보천련질보분석득도준학감정,포괄혈청전철단백、혈청β섬유단백원등.결론 통과단백질조학연구발현,혈청전철단백등단백질재애자병환자혈장중표체수평여치료과정중병독재량상관,타문가능시평개항병독치료료효적잠재생물표지물.
Objective To find specific biomarkers related to HAART treatment in plasma samples of AIDS patients for clinical therapeautic efficacy evaluation and guidance for the prognosis of HIV treatment.Method Plasma samples of AIDS patients were collected from Infectious Disease Department 1 of Shanghai Public Health Clinical Center in June of 2008 to February of 2009,including 11 successfully HAART treated cases (HIV load ＞ 50 copies/ml) and 11 unsuccessfully HAART treated cases (HIV load ＜50 copies/ml).Patients' age ranged from 22 to 63.Plasma samples were treated by Bio-rad AurumTM Serum Protein Mini Kit to remove high abundant proteins:albumin and immunoglobulin were removed.The treated plasma proteins were separated by two-dimensional electrophoresis and analyzed by electrophoretogram using Imagemaster software to find differentially-expressed proteins related to therapeutic efficacy.After digestion by trypsin,the differentially-expressed proteins were identified by online reversed-phase nano-flow liquid chromatography coupled with electrospray ionization ion trap mass spectrometry.Results Low abundant proteins were efficiently enriched after the AurumTM Serum Protein Mini Kit treatment.Six differentially-expressed proteins were detected while comparing successfully and unsuccessfully HAART treated group.These proteins were accurately identified by tandem Mass spectrometry (MS), including serum transferrin, serum β-fibrinogen, etc.Conclusions Our proteomic research revealed that the differentially-expressed proteins such as transferrin,which is related to plasma virus loading in AIDS patients in the process of treatment,might be potential biomarkers evaluating HAART therapeutic efficacy.
