Meijer A, van der Vliet J A, Schouls L M, de Vries A, Roholl P J, Ossewaarde J M
Research Laboratory for Infectious Diseases, National Institute of Public Health, Bilthoven, The Netherlands.
Res Microbiol. 1998 Sep;149(8):577-83. doi: 10.1016/s0923-2508(99)80005-9.
A procedure in which the "Invitrogen Easy-DNA" kit was followed by a silica-based method for the isolation of DNA was developed for extraction of PCR-inhibitor-free DNA from up to 300 mg of human vessel wall tissue. Optimally designed PCR assays were developed for the detection of at least one infected cell in this amount of tissue. Details of the procedure are given for the detection of DNA of Chlamydia pneumoniae, cytomegalovirus and herpes simplex virus type 1 and type 2 in human vessel walls. The procedure can serve as a reference method or as a gold standard when a high-performance method is needed.
我们开发了一种方法，先用“Invitrogen Easy-DNA”试剂盒，再采用基于硅胶的方法从多达300毫克的人体血管壁组织中提取无PCR抑制剂的DNA。针对在该组织量中检测至少一个感染细胞，开发了优化设计的PCR检测方法。文中给出了在人体血管壁中检测肺炎衣原体、巨细胞病毒以及单纯疱疹病毒1型和2型DNA的详细步骤。当需要一种高性能方法时，该方法可作为参考方法或金标准。
