Frank T S, Cook S M, Del Buono E A, Wilson M D
Department of Pathology, University of Michigan School of Medicine and Hospitals, Ann Arbor.
Mod Pathol. 1992 Jul;5(4):449-54.
Using a simplified procedure, we have extracted DNA from unstained paraffin sections of needle biopsies of kidney and liver transplants and identified the presence of CMV using the polymerase chain reaction. This method utilizes oligonucleotide primers for two genes shown to be specific for cytomegalovirus (CMV) as well as an internal control gene (hemoglobin) in a single reaction. Utilizing nested PCR amplification with agarose gel electrophoresis, CMV can be detected without radioisotopes to a level of sensitivity equivalent to one one-hundredth of a cytomegalic virocyte per cm2 of a 3-microM paraffin section. This method is applicable to situations where only scarce paraffin-embedded tissue is available.
我们采用一种简化程序，从肾脏和肝脏移植针吸活检的未染色石蜡切片中提取了DNA，并使用聚合酶链反应鉴定了巨细胞病毒（CMV）的存在。该方法在单一反应中利用针对两个已证明对巨细胞病毒（CMV）特异的基因以及一个内部对照基因（血红蛋白）的寡核苷酸引物。利用嵌套式聚合酶链反应扩增和琼脂糖凝胶电泳，无需放射性同位素即可检测到CMV，其灵敏度相当于每平方厘米3微米石蜡切片中一个巨细胞病毒细胞的百分之一。该方法适用于仅能获得少量石蜡包埋组织的情况。
