中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2010年
5期
389-391
,共3页
方丽娟%来乐祥%乐奕全%任玲君
方麗娟%來樂祥%樂奕全%任玲君
방려연%래악상%악혁전%임령군
肝炎病毒,乙型%突变%寡核苷酸序列分析
肝炎病毒,乙型%突變%寡覈苷痠序列分析
간염병독,을형%돌변%과핵감산서렬분석
Hepatitis B Virus%Mutation%Oligonucleotide array sequence analysis
目的 建立检测乙型肝炎病毒核心启动子及前C区突变基因芯片并探讨其临床应用的价值.方法 设计并合成针对核心启动子1762/1764、1814及前C区1896位点突变的特异性探针,制备寡核苷酸芯片.采用不对称PCR对该区域进行扩增,扩增产物与芯片杂交后分析结果,并评价该方法的特异性、灵敏度.采用该方法检测138例HBV DNA阳性血清标本.结果 该方法能够特异地检测乙型肝炎病毒核心启动子1762/1764、1814及前C区1896位点,灵敏度达1×101拷贝/μl.138例HBV DNA阳性血清标本中,T1762/A1764突变40例(28.99%),C1814突变11例(7.97%),A1896突变16例(11.59%).前C区A1896突变在高拷贝组中明显高于低拷贝组(P＜0.01).结论 本研究建立的基因芯片能够准确同时检测HBVV核心启动子区突变和前C区突变,前C区A1896突变可能与HBV复制状态有关.
目的 建立檢測乙型肝炎病毒覈心啟動子及前C區突變基因芯片併探討其臨床應用的價值.方法 設計併閤成針對覈心啟動子1762/1764、1814及前C區1896位點突變的特異性探針,製備寡覈苷痠芯片.採用不對稱PCR對該區域進行擴增,擴增產物與芯片雜交後分析結果,併評價該方法的特異性、靈敏度.採用該方法檢測138例HBV DNA暘性血清標本.結果 該方法能夠特異地檢測乙型肝炎病毒覈心啟動子1762/1764、1814及前C區1896位點,靈敏度達1×101拷貝/μl.138例HBV DNA暘性血清標本中,T1762/A1764突變40例(28.99%),C1814突變11例(7.97%),A1896突變16例(11.59%).前C區A1896突變在高拷貝組中明顯高于低拷貝組(P＜0.01).結論 本研究建立的基因芯片能夠準確同時檢測HBVV覈心啟動子區突變和前C區突變,前C區A1896突變可能與HBV複製狀態有關.
목적 건립검측을형간염병독핵심계동자급전C구돌변기인심편병탐토기림상응용적개치.방법 설계병합성침대핵심계동자1762/1764、1814급전C구1896위점돌변적특이성탐침,제비과핵감산심편.채용불대칭PCR대해구역진행확증,확증산물여심편잡교후분석결과,병평개해방법적특이성、령민도.채용해방법검측138례HBV DNA양성혈청표본.결과 해방법능구특이지검측을형간염병독핵심계동자1762/1764、1814급전C구1896위점,령민도체1×101고패/μl.138례HBV DNA양성혈청표본중,T1762/A1764돌변40례(28.99%),C1814돌변11례(7.97%),A1896돌변16례(11.59%).전C구A1896돌변재고고패조중명현고우저고패조(P＜0.01).결론 본연구건립적기인심편능구준학동시검측HBVV핵심계동자구돌변화전C구돌변,전C구A1896돌변가능여HBV복제상태유관.
Objective To develop a sensitive and specific microarray for detecting mutations of HBV pre-core/core and basic core promoter regions in the clinic. Methods Site-specific oligonucleotide probes were designed and immobilized to microarray slides and hybridized to HBV gene fragments amplified with specific biotin-labeled primer using asymmetrical PCR. The specificity and sensitivity of the method were estimated. And the microarray was applied to detect 138 clinical serum samples with HBV-DNA.Results The mutations of HBV pre-core/core and basic core promoter regions can be specifically detected using the microarray, and the sensitivity was 1 × 101 copies/μl. Among 138 samples,40 samples had T1762/A1764 mutation, 1 lsamples had C1814 mutation, and 16 samples had A1896 mutation. The A1896 mutation rate in high HBV-DNA load group was significantly higher than that in low HBV-DNA load group (P ＜ 0. 01 ). Conclusion An DNA microarray assay was successfully established to detect the mutations in HBV pre-core/core and basic core promoter regions. The A1896 mutation in Pre-core/core region maybe involve in duplication of HBV.
