Wilkens L, Werner M, Nolte M, Wasielewski R V, Verhagen W, Flik J, Klempnauer J, Georgii A
Pathologisches Institute, Hochschule Hannover, Germany.
Diagn Mol Pathol. 1994 Sep;3(3):156-62. doi: 10.1097/00019606-199409000-00004.
The possibility of detecting cytomegalovirus (CMV) in formalin-fixed tissues by polymerase chain reaction (PCR) was evaluated in necroposies from lung tissues in a total of 24 patients who either had received organ transplants or were immunocompromised. PCR using two different pairs of primers for amplification of the major immediate early antigen of CMV was performed on fresh tissues and tissues fixed for 24, 48, and 72 h in neutral buffered formalin and compared to immunohistochemistry (IHC) and in situ hybridization (ISH). The fresh tissues of nine patients with serological evidence for acute CMV infection were all positive for CMV by PCR. After formalin fixation, the majority of the patients failed to show distinct signals with one or both pairs of primers as measured by densitometry. In contrast to this, fresh tissues of 15 patients without signs of an acute CMV infection were found either negative or weakly positive by PCR. Using IHC or ISH, positive results were observed only in five of nine and four of nine patients with acute CMV infection, respectively. These data demonstrate that, if only formalin-fixed tissue is available, PCR for CMV detection should be performed using two pairs of primers and should be supported by IHC.
通过聚合酶链反应（PCR）在福尔马林固定组织中检测巨细胞病毒（CMV）的可能性，在总共24例接受器官移植或免疫功能低下患者的肺组织尸检中进行了评估。使用两对不同引物扩增CMV主要即刻早期抗原的PCR，在新鲜组织以及在中性缓冲福尔马林中固定24、48和72小时的组织上进行，并与免疫组织化学（IHC）和原位杂交（ISH）进行比较。9例有急性CMV感染血清学证据患者的新鲜组织，通过PCR检测CMV均呈阳性。福尔马林固定后，通过光密度测定法，大多数患者用一对或两对引物均未显示明显信号。与此相反，15例无急性CMV感染迹象患者的新鲜组织，通过PCR检测发现为阴性或弱阳性。使用IHC或ISH，分别仅在9例急性CMV感染患者中的5例和4例中观察到阳性结果。这些数据表明，如果仅有福尔马林固定组织可用，CMV检测的PCR应使用两对引物进行，并应以IHC作为辅助。
