Silva R F
U.S. Department of Agriculture, Agricultural Research Service, East Lansing, Michigan 48823.
Avian Dis. 1992 Jul-Sep;36(3):521-8.
There are no simple, direct methods to reliably distinguish oncogenic serotype 1 Marek's disease viruses (MDVs) from their attenuated variants. The present study was an attempt to apply polymerase chain reaction (PCR) to develop a rapid and sensitive assay for the presence of the MDV genome. PCR oligos were chosen to flank the 132-base-pair tandem direct repeats in the serotype 1 MDV genome. The PCR reaction was specific for serotype 1 MDVs, amplifying fragments corresponding to one to three copies of the tandem repeats present in Md11/8, JM/102W, and GA viruses. A high-molecular-weight DNA smear was observed when the DNA from an attenuated Md11/100 was PCR-amplified. Use of the PCR technique allowed the detection of two copies of the 132-base-pair repeat in the DNA extracted from MDV-induced lymphomas removed from two chickens. No DNA was amplified from the DNA extracted from lymphomas induced by either an avian leukosis virus (RAV-1) or reticuloendotheliosis virus (chick syncytial virus).
目前尚无简单、直接的方法能可靠地区分致瘤性1型马立克氏病病毒(MDV)与其减毒变体。本研究试图应用聚合酶链反应(PCR)来开发一种快速、灵敏的检测MDV基因组存在的方法。选择PCR寡核苷酸位于1型MDV基因组中132个碱基对的串联直接重复序列两侧。该PCR反应对1型MDV具有特异性,可扩增与Md11/8、JM/102W和GA病毒中存在的一至三个串联重复拷贝相对应的片段。当对减毒的Md11/100的DNA进行PCR扩增时,观察到高分子量DNA拖尾现象。使用PCR技术可检测到从两只鸡身上取出的MDV诱导淋巴瘤中提取的DNA中有两个132个碱基对重复序列的拷贝。从禽白血病病毒(RAV-1)或网状内皮组织增殖病病毒(鸡合胞体病毒)诱导的淋巴瘤中提取的DNA未扩增出任何DNA。