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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of Toxoplasma ELITe MGB Real-Time PCR Assay for Diagnosis of Toxoplasmosis
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Evaluation of Toxoplasma ELITe MGB Real-Time PCR Assay for Diagnosis of Toxoplasmosis

机译：弓形虫ELITe MGB实时PCR检测对弓形虫病诊断的评估

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Molecular diagnosis of congenital toxoplasmosis or disseminated toxoplasmosis is based mainly on PCR. The repeated DNA element rep529 has become the main DNA target used in most PCR methods, whether laboratory developed or commercial. In this multicenter study, we evaluated the Toxoplasma ELITe MGB (Elitech) commercial kit by comparison with three reference quantitative PCR assays (RAs) used routinely in three proficient laboratories of the French National Reference Center for Toxoplasmosis network, using Toxoplasma calibrated suspensions diluted to obtain a range of concentrations from 0.1 to 10,000 parasites/ml. These suspensions were extracted with either the DNA extraction kit (EXTRAblood; Elitech) recommended by the manufacturer or the QIAamp DNA minikit (Qiagen). The Toxoplasma ELITe MGB assay was also evaluated on a panel of 128 clinical samples, including 56 amniotic fluid samples, 55 placenta samples, and various other samples, of which 95 originated from patients with proven toxoplasmosis. The ELITe MGB assay amplified low-concentration replicates (<10 parasites/ml) of calibrated suspensions less frequently than the RAs of 2/3 laboratories. Additionally, the combination of EXTRAblood and Toxoplasma ELITe MGB yielded poorer sensitivity than the combination of QIAamp DNA minikit and ELITe MGB for low parasite concentrations (P < 0.001 for 1 parasite/ml). On clinical samples, the sensitivity and the specificity of the commercial assay were 89% and 100%, respectively. The sensitivity ranged from 79% (placenta samples) to 100% (amniotic fluid samples). Overall, this study shows that the Toxoplasma ELITe MGB assay is suitable for the diagnosis of toxoplasmosis from non-cell-rich or non-hemoglobin-rich samples and that the EXTRAblood kit is not optimal.
机译：先天性弓形虫病或弥散性弓形虫病的分子诊断主要基于PCR。重复的DNA元件 rep529 已成为大多数PCR方法中使用的主要DNA靶标，无论是实验室开发的还是商业化的。在此多中心研究中，我们通过与法国国家弓形虫病参考中心网络的三个熟练实验室中常规使用的三种参考定量PCR分析（RA）进行比较，对弓形虫ELITe MGB（Elitech）商业试剂盒进行了评估，并使用稀释后得到的弓形虫校正悬浮液浓度范围从0.1到10,000寄生虫/毫升。用制造商推荐的DNA提取试剂盒（EXTRAblood; Elitech）或QIAamp DNA minikit（Qiagen）提取这些悬浮液。弓形虫ELITe MGB分析还通过一组128个临床样品进行评估，包括56个羊水样品，55个胎盘样品和其他各种样品，其中95个来自已证明弓形虫病的患者。与2/3实验室的RA相比，ELITe MGB测定法可以更频繁地对校准悬浮液进行低浓度重复测定（<10寄生虫/ ml）。此外，对于低寄生虫浓度（ P <0.001，每1个寄生虫/ ml），EXTRAblood和弓形虫ELITe MGB的组合产生的敏感性比QIAamp DNA minikit和ELITe MGB的组合差。在临床样品上，商业检测的灵敏度和特异性分别为89％和100％。灵敏度范围从79％（胎盘样本）到100％（羊水样本）。总体而言，这项研究表明，弓形虫ELITe MGB检测法适合于从非富含细胞或非富含血红蛋白的样品中诊断弓形虫病，并且EXTRAblood试剂盒不是最佳的。

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