Donoso Mantke Oliver, Nitsche Andreas, Meyer Rudolf, Klingel Karin, Niedrig Matthias
Robert Koch-Institut, Nordufer 20, D-13353 Berlin, Germany.
J Clin Virol. 2004 Sep;31(1):32-9. doi: 10.1016/j.jcv.2003.12.013.
Parvovirus B19 (PVB19) is an erythrovirus causing diverse clinical manifestations ranging from asymptomatic or mild to more severe outcomes, dependent on the haematological and/or immunological status of the host. Reports of PVB19 infection as a causative agent of paediatric or adult inflammatory cardiac diseases, or of cardiac transplant rejection are rare.
To identify PVB19 and other cardiotropic viruses in the myocardium of heart transplant (HTx) recipients and multi-organ donors (MOD). Furthermore, to assess the prevalence of cardiotropic viral infection in inflammatory heart disease.
Heart tissue samples from 110 explants were analysed for PVB19 using primers and a 5'-nuclease probe designed to amplify a 160-basepair PCR product from the VP1/NS1 gene region. Samples tested included those obtained from patients undergoing HTx or from MODs. The findings were correlated with clinical course, histologic analysis and serologic testing. Confirmation of the positive PCR-results was done by sequencing and in situ hybridisation.
The new assay described here allows precise quantitation of viral load over 7 orders of magnitude (10(6) to 10(0)IU/assay). Measurable amounts of parvoviral genomes were detected in 4/56 (7%) explanted HTx-hearts and in 5/54 (9%) explanted MOD-hearts.
The newly developed real-time PCR is a rapid, sensitive and specific method to detect PVB19 infection in heart tissue. It will be a useful tool to address important questions regarding viral infections transmitted by transplantation, acute infections, relapses and complications involving late or chronic rejection.
细小病毒B19(PVB19)是一种红细胞病毒,根据宿主的血液学和/或免疫学状态,可引起从无症状或轻微到更严重后果的多种临床表现。关于PVB19感染作为小儿或成人炎症性心脏病的病原体或心脏移植排斥反应的报道很少。
在心脏移植(HTx)受者和多器官供体(MOD)的心肌中鉴定PVB19和其他嗜心性病毒。此外,评估炎症性心脏病中嗜心性病毒感染的患病率。
使用引物和5'核酸酶探针分析来自110个外植体的心脏组织样本中的PVB19,该探针设计用于从VP1/NS1基因区域扩增160个碱基对的PCR产物。测试的样本包括从接受HTx的患者或MOD获得的样本。研究结果与临床病程、组织学分析和血清学检测相关。通过测序和原位杂交对PCR阳性结果进行确认。
此处描述的新检测方法可对7个数量级(10⁶至10⁰IU/检测)的病毒载量进行精确定量。在4/56(7%)的HTx外植心脏和5/54(9%)的MOD外植心脏中检测到可测量数量的细小病毒基因组。
新开发的实时PCR是一种快速、灵敏且特异的检测心脏组织中PVB19感染的方法。它将成为解决有关移植传播的病毒感染、急性感染、复发以及涉及晚期或慢性排斥反应的并发症等重要问题的有用工具。