【摘要】 为了更准确、简便而又快速地了解乙型肝炎表面抗原（ＨＢｓＡｇ）阴性肝病患者的病因，建立了套式和免疫套式聚合酶链反应（ＰＣＲ）检测血清中乙型肝炎病毒（ＨＢＶ）ＤＮＡ的技术，结合丙型肝炎病毒（ＨＣＶ）感染指标的检测，对ＨＢｓＡｇ阴性肝病患者的病因进行了研究。发现套式ＰＣＲ能将单次ＰＣＲ的敏感性稳定地提高１０００倍；免疫套式ＰＣＲ可检测到０．１～０．０１ｐｇ／Ｌ水平。检测ＨＢｓＡｇ阴性肝硬化２２例（Ａ组）、ＨＢｓＡｇ阴性慢性肝炎１３例（Ｂ组）、ＨＢｓＡｇ阴性和乙型肝炎病毒核心抗体（抗－ＨＢｃ）阳性正常对照组３０例（Ｃ组）及ＨＢｓＡｇ（＋）／ＨＢｅＡｇ（－）肝硬化患者１２例（Ｄ组），分别有４５．５％、３０．８％、１３．３％和１００％患者血清中ＨＢＶＤＮＡ阳性。ＨＢＶＤＮＡ在一些抗－ＨＢｓ（＋）肝病患者和所谓健康人的血清中也存在。Ａ、Ｂ两组检出有ＨＢＶ和（或）ＨＣＶ感染患者分别占８１．８％和５３．８％。提示套式和免疫套式ＰＣＲ是简便、快速而又高度敏感的检测方法；ＨＢＶ感染可能是引起ＨＢｓＡｇ阴性慢性肝病的重要原因，且ＨＢｓＡｇ阴性肝病病因大多与病毒感染有关；应该重新认识自然感染者血清中抗－ＨＢｓ的临床意义。更多还原

【Abstract】 Inordertofindoutrapidlythecausesoftheliverdiseasessuferedbypatientswithneg-ativehepatitisBsurfaceantigen(HBsAg),nestedpolymerasechainreaction(PCR)andmultipleclonedantibodycapturePCRtechniqueswereestablishedtoexamineserumhepatitisBvirus(HBV)DNA.ByusingbothtechniquesalongwiththeexaminationofhepatitisCvirus(HCV)infection,thecausesofchronicliverdiseaseswithnegativeHBsAgwerestudied.Itisfoundthatnested-PCRcanincreasethesensitivityofsinglePCRmorethan1000foldandmultipleclonedantibodycapture-PCRcandetectcon-centrationofHBVDNAaslowas0.1～0.01pg/L.HBVDNApositivepatientswerefoundin45.5%,30.8%,13.3%and100%respectivelyofthepatientssuferingfromlivercirhosiswithnegativeHBsAg(groupA,22cases),chronichepatitiswithnegativeHBsAg(groupB,13cases),normalsubjectswithnegativeHBsAgandpositivehepatitisBcoreantibody(HBcAb,groupC,30cases)andlivercirhosiswithpositiveHBsAgandnegativeHBeAg(groupD,12cases).HBVDNAcanbealsofoundintheserumofHBsAbpositivepatientsandsubjectssupposedtobehealthy.81.8%and53.8%ofthepatientswereinfectedwithHBVand/orHCVingroupAandgroupBrespectively.Altheseresultssuggestthatnested-PCRandmultipleclonedantibodycaputure-PCRarerapidandhighlysensitivemethodsforde-tectionofserumHBVDNA.HBVinfectionisanimportantcauseofchronicliverdiseasesinpatientswithnegativeHBsAg.ThecausesofmostoftheHBsAg-negativechronicliverdiseasesarerelatedwithinfectionofviruses.TheclinicalsignificanceofserumHBsAbinnaturalyinfectedpatientsshouldbere-considered.更多还原