中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
7期
665-669
,共5页
王垚%苏丽菊%李辉%刘彦虹%翟爱霞%考文萍%吴静%李文辉%胡云龙%钟照华%张凤民
王垚%囌麗菊%李輝%劉彥虹%翟愛霞%攷文萍%吳靜%李文輝%鬍雲龍%鐘照華%張鳳民
왕요%소려국%리휘%류언홍%적애하%고문평%오정%리문휘%호운룡%종조화%장봉민
噬菌体随机肽库%系统性红斑狼疮%血清
噬菌體隨機肽庫%繫統性紅斑狼瘡%血清
서균체수궤태고%계통성홍반랑창%혈청
Phage peptide library%Systemic lupus erythematosus%Sera
目的 筛选和鉴定与系统性红斑狼疮(systemic lupus erythematosus,SLE)患者血清特异性结合的噬菌体7肽,并分析其实际意义.方法 分别选取正常人及SLE患者血清各30例,先后用正常人混合血清及SLE患者混合血清作为筛选配基,对噬菌体随机7肽库进行亲和筛选、扩增,获得SLE血清特异性结合的噬菌体克隆,并用患者混合血清进行Dot-ELISA实验鉴定获得的噬菌体克隆,进而分别用SLE患者及正常人血清各12例进一步鉴定阳性噬菌体的混合克隆,确定阳性噬菌体克隆与个体血清之间的结合情况;并对最终鉴定的噬菌体克隆进行测序与比对分析.结果 筛选到与SLE患者混合血清特异性结合的阳性克隆12个;阳性噬菌体混合克隆与SLE患者个体血清反应阳性率明显高于其与正常人血清的反应率;序列分析显示阳性噬菌体克隆的抗原表位与大肠杆菌、沙门菌、人类免疫缺陷病毒(HIV)有一定的同源性,但与人类抗原表位无关.结论 噬菌体随机7肽库筛选出的SLE特异性多肽可能用于制备SLE诊断试剂,同时SLE患者血清中存在与病原体抗原表位结合的抗体成分,提示SLE可能与病原体感染有关.
目的 篩選和鑒定與繫統性紅斑狼瘡(systemic lupus erythematosus,SLE)患者血清特異性結閤的噬菌體7肽,併分析其實際意義.方法 分彆選取正常人及SLE患者血清各30例,先後用正常人混閤血清及SLE患者混閤血清作為篩選配基,對噬菌體隨機7肽庫進行親和篩選、擴增,穫得SLE血清特異性結閤的噬菌體剋隆,併用患者混閤血清進行Dot-ELISA實驗鑒定穫得的噬菌體剋隆,進而分彆用SLE患者及正常人血清各12例進一步鑒定暘性噬菌體的混閤剋隆,確定暘性噬菌體剋隆與箇體血清之間的結閤情況;併對最終鑒定的噬菌體剋隆進行測序與比對分析.結果 篩選到與SLE患者混閤血清特異性結閤的暘性剋隆12箇;暘性噬菌體混閤剋隆與SLE患者箇體血清反應暘性率明顯高于其與正常人血清的反應率;序列分析顯示暘性噬菌體剋隆的抗原錶位與大腸桿菌、沙門菌、人類免疫缺陷病毒(HIV)有一定的同源性,但與人類抗原錶位無關.結論 噬菌體隨機7肽庫篩選齣的SLE特異性多肽可能用于製備SLE診斷試劑,同時SLE患者血清中存在與病原體抗原錶位結閤的抗體成分,提示SLE可能與病原體感染有關.
목적 사선화감정여계통성홍반랑창(systemic lupus erythematosus,SLE)환자혈청특이성결합적서균체7태,병분석기실제의의.방법 분별선취정상인급SLE환자혈청각30례,선후용정상인혼합혈청급SLE환자혼합혈청작위사선배기,대서균체수궤7태고진행친화사선、확증,획득SLE혈청특이성결합적서균체극륭,병용환자혼합혈청진행Dot-ELISA실험감정획득적서균체극륭,진이분별용SLE환자급정상인혈청각12례진일보감정양성서균체적혼합극륭,학정양성서균체극륭여개체혈청지간적결합정황;병대최종감정적서균체극륭진행측서여비대분석.결과 사선도여SLE환자혼합혈청특이성결합적양성극륭12개;양성서균체혼합극륭여SLE환자개체혈청반응양성솔명현고우기여정상인혈청적반응솔;서렬분석현시양성서균체극륭적항원표위여대장간균、사문균、인류면역결함병독(HIV)유일정적동원성,단여인류항원표위무관.결론 서균체수궤7태고사선출적SLE특이성다태가능용우제비SLE진단시제,동시SLE환자혈청중존재여병원체항원표위결합적항체성분,제시SLE가능여병원체감염유관.
Objective To screen and identify the phage-display random 7 amino acid peptide specific to the systemic lupus erythematosus(SLE) and analyze its practical significance. Methods Using the phage random 7 peptide library screening, the SLE specific phage clones are obtained after binding with the mixture of sera from 30 SLE patients and 30 normal controls as ligand respectively. Then the Dot-ELISA is used to identify the SLE specific phage clones reactive to sera of the SLE patients and normal controls individually. Finally the identified phage-display random 7 amino acid peptides are sequenced and it's homology with the antigenic epitope of human being and other are also analyzed. Results Total 12 of the phage-display random 7 amino acid peptide are obtained by phage peptide library screening and the Dot-ELISA identification. Sequence analysis shows that the identified phage-display random 7 amino acid peptide epitope have homology with E. coli, Salmonella and human immunodeficiency virus, but not with that of human being. Conclusion SLE-specific peptides screened by phage random peptide library maybe used to diagnosis the SLE. Meanwhile, the antibodies in SLE patients which are combined with the Pathogen epitope, suggest that SLE maybe relate to pathogen infection.
