应用TaqMan聚合酶链反应定量检测人体体液中的弓形虫DNA
Quantitative detection of Toxoplasma gondii DNA in human body fluids by TaqMan polymerase chain reaction.
作者信息
Kupferschmidt O, Krüger D, Held T K, Ellerbrok H, Siegert W, Janitschke K
机构信息
Robert-Koch-Institut, Parasitologie/Mykologie, Nordufer 20, D-13353 Berlin, Germany.
出版信息
Clin Microbiol Infect. 2001 Mar;7(3):120-4. doi: 10.1046/j.1469-0691.2001.00224.x.
OBJECTIVE
A new quantitative polymerase chain reaction (real-time PCR) was designed to detect Toxoplasma DNA in human body fluid samples.
METHODS
Real-time fluorescence detection of amplification product formation on the basis of the TaqMan-System was established with Toxoplasma 18S rDNA as a target gene.
RESULTS
The method provides a high sensitivity comparable to conventional nested PCR procedures and generates quantitative data when detecting toxoplasmic DNA in human blood, cerebrospinal or amniotic fluid. Moreover, data were obtained investigating blood samples from an immunocompromised patient with reactivated toxoplasmosis after allogeneic bone marrow transplantation, monitoring the therapeutic effect.
CONCLUSIONS
The potential application of this method to detect Toxoplasma DNA in body fluids and to follow the development of parasitemia under therapy could be demonstrated.
目的
设计一种新的定量聚合酶链反应（实时荧光定量PCR）方法来检测人体体液样本中的弓形虫DNA。
方法
以弓形虫18S rDNA为靶基因，基于TaqMan系统建立扩增产物形成的实时荧光检测方法。
结果
该方法具有与传统巢式PCR相当的高灵敏度，在检测人血液、脑脊液或羊水弓形虫DNA时可产生定量数据。此外，通过对一名异基因骨髓移植后弓形虫病复发的免疫功能低下患者的血液样本进行研究，获得了数据以监测治疗效果。
结论
该方法在检测体液中弓形虫DNA以及监测治疗过程中虫血症发展方面的潜在应用得到了证实。
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