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[通过R基因实时荧光定量PCR检测评估血液中巨细胞病毒定量]

[Evaluation of cytomegalovirus quantification in blood by the R-gene real-time PCR test].

作者信息

Marque-Juillet S, Touzard A, Monnier S, Fernand-Laurent C, Therby A, Rigaudeau S, Harzic M

机构信息

Service de microbiologie, centre hospitalier de Versailles, 177, rue Versailles, 78150 Le Chesnay, France.

出版信息

Pathol Biol (Paris). 2010 Apr;58(2):162-5. doi: 10.1016/j.patbio.2009.07.029. Epub 2009 Oct 24.

Abstract

AIM OF THE STUDY

Diagnosing the presence of cytomegalovirus (CMV) in the blood of immunodepressed patients is often done by quantitative polymerase chain reaction (Q-PCR) even though the reference method remains the antigenemia pp65 (Ag-pp65) test.

OBJECTIVES

To define the predictive value of the Q-PCR in the diagnosis of CMV disease and assess treatment efficacy using the CMV R-gene test. To compare the Q-PCR results and feasibility with those of the Ag-pp65 test.

PATIENTS AND METHODS

The Q-PCR was performed in 34 whole blood samples (frozen at -80 degrees C until use) from five patients diagnosed with CMV disease, defined as the presence of clinical signs and Ag-pp65 in the nuclei of more than two cells. After extraction, viral DNA was quantified in each sample using the Q-PCR CMV R-gene kit according to the manufacturer's instructions. Immediately after blood was drawn, the Ag-pp65 test had been performed in 32 samples using CINAkit (Argene).

RESULTS

The 16 samples positive by the Ag-pp65 test were also positive by PCR; six samples negative by the Ag-pp65 test were positive by PCR; and the remaining 10 samples were negative by both techniques. During treatment, the two markers' kinetics were similar.

CONCLUSION

The CMV R-gene test has a predictive value as good as that of the Ag-pp65 test but is fast and easier to use. A prospective study with a greater number of patients is needed to define the prediction threshold for CMV disease.

摘要

研究目的

尽管参考方法仍是抗原血症pp65(Ag-pp65)检测,但免疫抑制患者血液中巨细胞病毒(CMV)的诊断通常采用定量聚合酶链反应(Q-PCR)。

目的

确定Q-PCR在CMV疾病诊断中的预测价值,并使用CMV R基因检测评估治疗效果。比较Q-PCR结果与Ag-pp65检测结果及可行性。

患者和方法

对5例诊断为CMV疾病(定义为存在临床体征且在两个以上细胞的细胞核中检测到Ag-pp65)患者的34份全血样本(在-80℃冷冻保存直至使用)进行Q-PCR检测。提取后,根据制造商说明,使用Q-PCR CMV R基因试剂盒对每个样本中的病毒DNA进行定量。采血后立即使用CINAkit(Argene)对32份样本进行Ag-pp65检测。

结果

Ag-pp65检测呈阳性的16份样本PCR检测也呈阳性;Ag-pp65检测呈阴性的6份样本PCR检测呈阳性;其余10份样本两种技术检测均为阴性。治疗期间,两种标志物的动态变化相似。

结论

CMV R基因检测的预测价值与Ag-pp65检测相当,但速度更快且使用更简便。需要进行更多患者的前瞻性研究以确定CMV疾病的预测阈值。

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