首页> 外文期刊>Journal of Clinical Microbiology >Comparison of LightCycler-Based PCR, COBAS Amplicor CMV Monitor, and pp65 Antigenemia Assays for Quantitative Measurement of Cytomegalovirus Viral Load in Peripheral Blood Specimens from Patients after Solid Organ Transplantation
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Comparison of LightCycler-Based PCR, COBAS Amplicor CMV Monitor, and pp65 Antigenemia Assays for Quantitative Measurement of Cytomegalovirus Viral Load in Peripheral Blood Specimens from Patients after Solid Organ Transplantation

机译:基于LightCycler的PCR,COBAS Amplicor CMV监测仪和pp65抗原血症测定法对定量测量固体器官移植患者外周血中巨细胞病毒载量的比较

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In order to evaluate the LightCycler-based PCR (LC-PCR) as a diagnostic assay technique, a classical pp65 antigenemia assay and the commercially available COBAS Amplicor CMV Monitor (CACM) assay were compared to the LC-PCR assay for the detection and quantitation of cytomegalovirus (CMV) load in 404 parallel specimens of peripheral blood from 66 patients after solid organ transplantation. A good correlation existed among these three assays (r ? 0.6, P < 0.0001). The LC-PCR assay was the most sensitive (54% of specimens positive) compared to the CACM (48.6%) and the pp65 antigenemia (26%) assays. The LC-PCR assay detected all samples found positive by using both the CMV pp65 antigenemia assay and the CACM assay. The LC-PCR also had the widest dynamic range (from 250 to 107 DNA copies/ml of plasma). No cross-reactions were found among CMV and Epstein-Barr virus, varicella-zoster virus, or herpes simplex virus in the LC-PCR by using amplification with specifically designed primer pairs. Precision, expressed as the coefficient of variation, was <3% with standard DNA from cell cultures and between 6.55 and 14.1% with clinical specimens in repeat LC-PCR runs. One run of the LC-PCR took half of the time required for the semiautomated CACM procedure. Because of its sensitivity, specificity, cost-effectiveness, and simplicity, the LC-PCR assay could replace the pp65 antigenemia and the CACM assays as the preferred technique for the surveillance, diagnosis, and monitoring of response of CMV diseases in high-risk populations.
机译:为了评估基于LightCycler的PCR(LC-PCR)作为诊断测定技术,将经典的pp65抗原血症测定和市售的COBAS Amplicor CMV Monitor(CACM)测定与LC-PCR测定进行了比较,以进行检测和定量实体器官移植后66例患者的404平行血液样本中巨细胞病毒(CMV)负荷的变化这三个测定之间存在良好的相关性( r ≤0.6, P <0.0001)。与CACM(48.6%)和pp65抗原血症(26%)分析相比,LC-PCR分析最为敏感(54%呈阳性)。 LC-PCR检测使用CMV pp65抗原血症检测和CACM检测检测到所有阳性的样品。 LC-PCR还具有最宽的动态范围(从250到10 7 DNA拷贝/ ml血浆)。通过使用专门设计的引物对进行扩增,LC-PCR中的CMV和Epstein-Barr病毒,水痘-带状疱疹病毒或单纯疱疹病毒之间未发现交叉反应。在重复LC-PCR运行中,用细胞培养物的标准DNA表示的精密度<3%,而使用临床标本的精确度在6.55%至14.1%之间。一次LC-PCR运行花费了半自动CACM程序所需时间的一半。由于其灵敏度,特异性,成本效益和简便性,LC-PCR测定法可替代pp65抗原血症和CACM测定法,作为监测,诊断和监测高危人群CMV疾病反应的首选技术。

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