Virology Laboratory, Hôpital Necker Enfants malades, APHP, Université Paris Descartes, Sorbonne Paris Cité-EA 36-20, Paris, France.
J Virol Methods. 2013 Nov;193(2):503-7. doi: 10.1016/j.jviromet.2013.07.024. Epub 2013 Jul 24.
In countries with limited resources, infants infected with HIV are highly exposed to CMV co-infection which probably represents a major risk factor for disease progression in this population. This study aimed to evaluate the performance of a low cost CMV DNA extraction method from DBS and the feasibility of its implementation in laboratories of 4 countries with limited resources. DNA was extracted from DBS with a cationic resin (chelex 100) and amplified with an "in house" real time CMV PCR. Dilutions of a quantified whole blood sample were spotted on paper to evaluate the 95% detection limit. A DBS quality control panel was analyzed in all laboratories. CMV PCR was compared between DBS and liquid whole blood (gold standard) in 2 populations: 418 transplanted patients and 59 infants infected with HIV (median age of 2 months). The CMV PCR 95% detection limit in DBS was 3.87 log10 copies/mL. Its positive and negative predictive values for CMV diagnosis in infants infected with HIV were 100% and 87.5% respectively. Quality control panels gave consistent qualitative results in all laboratories. This assay had high predictive values for CMV diagnosis in infants infected with HIV and its implementation in resource-limited countries with limited resources is feasible.
在资源有限的国家,感染 HIV 的婴儿高度暴露于 CMV 合并感染,这可能是该人群疾病进展的主要危险因素。本研究旨在评估从 DBS 中提取 CMV DNA 的低成本方法的性能,以及在资源有限的 4 个国家的实验室中实施该方法的可行性。使用阳离子树脂(chelex 100)从 DBS 中提取 DNA,并使用“内部”实时 CMV PCR 进行扩增。将定量全血样本的稀释液点在纸上,以评估 95%检测限。在所有实验室中分析 DBS 质量控制面板。在 2 个人群中比较 DBS 和全血(金标准)之间的 CMV PCR:418 名移植患者和 59 名感染 HIV 的婴儿(中位年龄为 2 个月)。DBS 中 CMV PCR 的 95%检测限为 3.87 log10 拷贝/mL。其对感染 HIV 的婴儿 CMV 诊断的阳性和阴性预测值分别为 100%和 87.5%。质量控制面板在所有实验室中给出一致的定性结果。该检测方法对感染 HIV 的婴儿 CMV 诊断具有较高的预测值,在资源有限的国家实施具有可行性。