本发明提供可用于诊断和监测治疗进展的、对干血斑检测HIV‑1病毒载量的新的和非显而易见的改进。
背景
人类免疫缺陷病毒(HIV)是获得性免疫缺陷综合征(AIDS)的病原体。(Barre-Sinoussi F, Chermann JC, Rey F等,从面临获得性免疫缺陷综合征(AIDS)风险的患者分离T-亲淋巴逆转录病毒(Isolation of a T-lymphotropic retrovirus from a patientat risk for acquired immune deficiency syndrome (AIDS)). Science 1983, 220:868-71; Popovic M, Sarngadharan MG, Read E等,来自AIDS和前-AIDS患者的细胞病变的逆转录病毒(HTLV-I)的检测、分离和连续生产(Detection, isolation and continuousproduction of cytopathic retroviruses (HTLV-I) from patients with AIDS andpre-AIDS). Science 1984, 224:497-500; Gallo RC, Salahuddin SZ, Popovic M等,从AIDS和面临AIDS风险的患者频繁检测和分离细胞病变的逆转录病毒(HTLV-I) (Frequentdetection and isolation of cytopathic retroviruses (HTLV-I) from patientswith AIDS and at risk for AIDS). Science 1984, 224:500-3)。它可以通过性接触、暴露于受感染的血液或血液制品,或从被感染的母亲到胎儿传播。(Curran JW, Jaffe HW,Hardy AM等,HIV感染和AIDS在美国的流行病学(Epidemiology of HIV infection andAIDS in the United States). Science 1988, 239:610-16)。急性HIV综合征,以流感样症状为特征,在开始感染后发展3-5周并与高水平的病毒血症相关。(Daar ES, Moudgil T,Meyer RD, Ho DD. 原发性人类免疫缺陷病毒1型感染患者中短暂的高水平病毒血症(Transient high levels of viremia in patients with primary humanimmunodeficiency virus type 1 infection). New Engl J Med 1991, 324:961-4;Clark SJ, Saag MS, Decker WD. 有症状的原发性HIV-1感染患者血浆中高滴度的细胞病变的病毒(High titers of cytopathic virus in plasma of patients withsymptomatic primary HIV-1 infection). New Engl J Med 1991, 324:954-60)。在症状发作的4-6周内,HIV特异性免疫应答是可检测的。(Albert J, Abrahamsson B, Nagy K等,在原发性HIV-1感染和随后的抗自体血清中和的病毒变种出现后,分离株-特异性中和抗体的快速发展(Rapid development of isolate-specific neutralizing antibodiesafter primary HIV-1 infection and consequent emergence of virus variantswhich resist neutralization by autologous sera). AIDS 1990, 4:107-12;Horsburgh CR Jr, Ou CY, Jason J等,在抗体检测前,人类免疫缺陷病毒感染的持续时间(Duration of human immunodeficiency virus infection before detection ofantibody). Lancet 1989, 334:637-40)。在血清转化后,在外周血中的病毒载量下降并且大多数患者进入可持续数年的无症状阶段。(Pantaleo G, Graziosi C, Fauci AS. 人类免疫缺陷病毒(HIV)感染的免疫发病机理的新概念(New concepts in theimmunopathogenesis of human immunodeficiency virus (HIV) infection). New Engl J Med 1993, 328:327-35)。外周血中HIV水平的定量测量大大有助于对HIV感染的发病机理的理解(Ho DD, Neumann AU, Perelson AS等,HIV-1感染中血浆病毒体和CD4淋巴细胞的快速周转(Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1infection). Nature 1995, 373:123-6; Wei X, Ghosh SK, Taylor ME等,人类免疫缺陷病毒1型感染的病毒动力学(Viral dynamics in human immunodeficiency virus type 1infection). Nature 1995, 373:117-22)并且已被证明是HIV感染个体的预后与管理中的一个重要参数。(Mellors JW, Rinaldo CR JR, Gupta P等,通过血浆中病毒的量预测的HIV-1感染的预后(Prognosis in HIV-1 infection predicted by the quantity ofvirus in plasma). Science 1996, 272:1167-70; Mellors JW, Munoz A, Giorgi JV等,作为HIV-1感染的预后标记物的血浆病毒载量和CD4+淋巴细胞(Plasma viral loadand CD4+ lymphocytes as prognostic markers of HIV-1 infection). Ann Intern Med 1997, 126(12):946-54; Chene G, Sterne JA, May M等,在开始有效的抗逆转录病毒疗法的HIV-1感染患者中初始应答的预后意义:前瞻性研究的分析(Prognosticimportance of initial response in HIV-1 infected patients starting potentantiretroviral therapy: analysis of prospective studies). Lancet 2003, 362:679-86; Egger M, May M, Chene G等,开始高活性抗逆转录病毒药物疗法的HIV-1感染患者的预后:一项前瞻性研究的合作分析(Prognosis of HIV-1 infected drug patientsstarting highly active antiretroviral therapy: a collaborative analysis ofprospective studies). Lancet 2002, 360:119-29; Wood E, Hogg RS, Yip B等,血浆人类免疫缺陷病毒1型RNA的较高基线水平与在三联药抗逆转录病毒疗法开始后死亡率增加有关(Higher baseline levels of plasma human immunodeficiency virus type 1RNA are associated with increased mortality after initiation of triple-drugantiretroviral therapy). J Infect Dis 2003, 188:1421-5; 美国卫生和人类服务部(US Department of Health and Human Services). 2004年HIV-1感染的成人和青少年中抗逆转录病毒药物的使用指南(2004 guidelines for the use of antiretroviralagents in HIV-1 infected adults and adolescents). 在:AIDSinfo.nih.gov/guidelines在线获得)。关于抗逆转录病毒疗法的开始或变化的决定通过监测血浆HIV RNA水平(病毒载量)、CD4+ T细胞计数,和患者的临床病症来指导。美国卫生和人类服务部(USDepartment of Health and Human Services). 2004年HIV-1感染的成人和青少年中抗逆转录病毒药物的使用指南(2004 guidelines for the use of antiretroviral agentsin HIV-1 infected adults and adolescents). 在:AIDSinfo.nih.gov/guidelines在线获得; Yeni PG, Hammer SM, Hirsch MS等,成人HIV感染的治疗(Treatment for AdultHIV Infection). 2004年国际AIDS协会-美国小组的建议(2004 Recommendations of theInternational AIDS Society-USA Panel). JAMA 2004, 292:251-65)。抗逆转录病毒疗法的目的是减少血浆中的HIV病毒至低于可利用的病毒载量试验的可检测水平。(美国卫生和人类服务部(US Department of Health and Human Services). 2004年HIV-1感染的成人和青少年中抗逆转录病毒药物的使用指南(2004 guidelines for the use ofantiretroviral agents in HIV-1 infected adults and adolescents). 在:AIDSinfo.nih.gov/guidelines在线获得; AS, Essunger P, Cao Y等,在组合疗法期间HIV-1感染的隔室的衰减特性(Decay characteristics of HIV-1 infectedcompartments during combination therapy). Nature 1997, 387(6629):188-91)。血浆中的HIV RNA水平可通过现有技术程序,用核酸扩增或信号扩增技术量化。(Mulder J,McKinney N, Christopher C等,用于血浆中人类免疫缺陷病毒1型RNA的定量的快速和简单的PCR测定:急性逆转录病毒感染的应用(Rapid and simple PCR assay forquantitation of human immunodeficiency virus type 1 RNA in plasma:application to acute retroviral infection). J Clin Microbiol 1994, 32:292-300; Dewar RL, Highbarger HC, Sarmiento MD等,分支DNA信号扩增以监测人血浆中的人类免疫缺陷病毒1型负担的应用(Application of branched DNA signalamplification to monitor human immunodeficiency virus type 1 burden in humanplasma). J Inf Diseases 1994, 170:1172-9; Van Gemen B, Kievits T, Schukkink R等,在HIV-1原发性感染期间,使用NASBA™量化血浆中的HIV-1 RNA (Quantification ofHIV-1 RNA in plasma using NASBA™ during HIV-1 primary infection). J Virol Methods 1993, 43:177-87)。