Real-time PCR targeting a 529-bp repeat element for diagnosis of toxoplasmosis
Real-time PCR targeting a 529-bp repeat element for diagnosis of toxoplasmosis
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DOI：
10.1111/j.1469-0691.2005.01332.x
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发表时间：
2006-02-01
影响因子：
14.2
通讯作者：
Evengård, B
中科院分区：
文献类型：
--
作者：
Edvinsson, B;Lappalainen, M;Evengård, B
在移植免疫功能低下的患者中，敏感和快速地检测弓形虫感染是获得良好预后的关键。目前用于检测弓形虫感染的DNA片段有两个，即B1基因和存在于200-300个拷贝/基因组中的529个核苷酸重复序列。这项研究调查了当弓形虫DNA浓度较低时，靶向529-BP重复元件是否比靶向B1基因(35个拷贝)具有更好的敏感性和准确性。结果表明，529bp重复序列的检测提高了诊断的敏感性和准确性。添加内部扩增对照不会影响聚合酶链式反应的性能，并且对于监测LightCycler仪器中非特异性DNA对聚合酶链式反应的抑制是有用的。实时聚合酶链式反应被成功地应用于临床，监测患有弓形虫病的移植受者血液中的寄生虫血症。
Sensitive and rapid detection of infection with Toxoplasma gondii in transplanted immunocompromised patients is crucial for a good prognosis. Two DNA fragments are used currently for detecting T. gondii infection by PCR, i.e., the B1 gene and a 529-bp repeat element that exists in 200-300 copies/genome. This study investigated whether targeting the 529-bp repeat element gives better sensitivity and accuracy than can be obtained when targeting the B1 gene (35 copies) when concentrations of T. gondii DNA are low. The results demonstrated that detection of the 529-bp repeat element increased diagnostic sensitivity and accuracy. Addition of an internal amplification control did not affect the PCR performance and was useful in order to monitor PCR inhibition by non-specific DNA in the LightCycler instrument. The real-time PCR was used successfully in a clinical context to monitor parasitaemia in the blood of a transplant recipient suffering from toxoplasmosis.