使用定量竞争性聚合酶链反应测量患有淋巴增生性疾病的儿科移植患者外周血中爱泼斯坦-巴尔病毒基因组载量。
Use of quantitative competitive PCR to measure Epstein-Barr virus genome load in the peripheral blood of pediatric transplant patients with lymphoproliferative disorders.
作者信息
Rowe D T, Qu L, Reyes J, Jabbour N, Yunis E, Putnam P, Todo S, Green M
机构信息
Department of Infectious Diseases and Microbiology, Graduate School of Public Health, The Childrens Hospital of Pittsburgh, Pennsylvania 15261, USA.
出版信息
J Clin Microbiol. 1997 Jun;35(6):1612-5. doi: 10.1128/jcm.35.6.1612-1615.1997.
A quantitative competitive PCR (QC-PCR) assay for Epstein-Barr virus (EBV) has been developed to provide accurate measurement of EBV genome load in pediatric transplant recipients at risk for developing posttransplant lymphoproliferative disorder (PTLD). The assay quantifies between 8 and 5,000 copies of the EBV genome in 10(5) lymphocytes after a 30-cycle amplification reaction. For 14 pediatric patients diagnosed with PTLD, the median EBV genome load was 4,000, and 13 of the 14 patients had values of >500 copies per 10(5) lymphocytes. Only 3 of 12 control transplant recipients not diagnosed with PTLD had detectable viral genome loads (median value, 40). This median was calculated by using the highest value obtained by PCR testing on each of these patients posttransplantation. PCR values of >500 copies per 10(5) lymphocytes appear to correlate with a diagnosis of PTLD. By a modified protocol, the EBV genome copy number in latently infected adults was estimated to be <0.1 copy per 10(5) lymphocytes.
已开发出一种用于检测爱泼斯坦-巴尔病毒（EBV）的定量竞争性聚合酶链反应（QC-PCR）检测方法，以准确测量有发生移植后淋巴细胞增生性疾病（PTLD）风险的儿科移植受者体内的EBV基因组载量。该检测方法在30个循环的扩增反应后，可对10⁵个淋巴细胞中的8至5000个EBV基因组拷贝进行定量。对于14例诊断为PTLD的儿科患者，EBV基因组载量中位数为4000，14例患者中有13例每10⁵个淋巴细胞的值>500拷贝。12例未诊断为PTLD的对照移植受者中只有3例可检测到病毒基因组载量（中位数为40）。该中位数是通过对这些患者移植后进行的每次PCR检测获得的最高值计算得出的。每10⁵个淋巴细胞的PCR值>500拷贝似乎与PTLD的诊断相关。通过改进方案，估计潜伏感染成年人的EBV基因组拷贝数为每10⁵个淋巴细胞<0.1拷贝。
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