de Vries Jutte J C, Barbi Maria, Binda Sandro, Claas Eric C J
Department of Medical Microbiology, Clinical Microbiological Laboratory, Leiden University Medical Center (LUMC), Leiden, The Netherlands.
Methods Mol Biol. 2012;903:169-75. doi: 10.1007/978-1-61779-937-2_10.
Viral DNA detection in dried blood spotted on filter paper, dried blood spots (DBS), is valuable in the diagnosis of viral infections, with at the moment congenital cytomegalovirus (CMV) being the most common application. CMV detection in clinical samples taken within the first 2-3 weeks after birth differentiates congenital CMV infection from the in general harmless postnatal acquired cytomegalovirus infection. DBS render the possibility to diagnose congenital CMV infection retrospectively, e.g., when late-onset hearing loss, the most frequently encountered symptom of congenital CMV infection, becomes manifest. Additionally, CMV DNA detection in DBS can be of usage in recently advocated newborn screening on congenital CMV infection. The procedure of CMV DNA detection in DBS consists of two separate steps: (1) DNA extraction from the DBS, followed by (2) CMV DNA amplification. Here, we describe two efficient methods for the extraction of DNA from DBS. Sensitivity, specificity, and applicability of the methods for high-throughput usage are discussed.
在滤纸上的干血斑（DBS）中进行病毒DNA检测，对于病毒感染的诊断具有重要价值，目前先天性巨细胞病毒（CMV）感染是其最常见的应用。在出生后最初2 - 3周内采集的临床样本中检测CMV，可将先天性CMV感染与通常无害的出生后获得性巨细胞病毒感染区分开来。干血斑使得回顾性诊断先天性CMV感染成为可能，例如，当先天性CMV感染最常见的症状——迟发性听力损失出现时。此外，干血斑中的CMV DNA检测可用于最近提倡的先天性CMV感染新生儿筛查。干血斑中CMV DNA检测程序包括两个独立步骤：（1）从干血斑中提取DNA，接着（2）进行CMV DNA扩增。在此，我们描述两种从干血斑中提取DNA的有效方法。并讨论了这些方法用于高通量检测时的灵敏度、特异性和适用性。
