Loparev V N, Massung R F, Esposito J J, Meyer H
Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, US Department of Health and Human Services, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 2001 Jan;39(1):94-100. doi: 10.1128/JCM.39.1.94-100.2001.
A restriction fragment length polymorphism (RFLP) assay was developed to identify and differentiate Old World, African-Eurasian orthopoxviruses (OPV): variola, vaccinia, cowpox, monkeypox, camelpox, ectromelia, and taterapox viruses. The test uses amplicons produced from virus genome DNA by PCR with a consensus primer pair designed from sequences determined for the cytokine response modifier B (crmB) gene of 43 different OPV strains of known taxonomic origin. The primer pair amplified a single specific product from each of the 115 OPV samples tested. Size-specific amplicons identified and differentiated ectromelia and vaccinia virus strains, which contain a truncated crmB gene, and enabled their differentiation from other OPV species. Restriction digests of amplified products allowed the identification and differentiation of variola, monkeypox, camelpox, vaccinia, and cowpox virus species and strains.
开发了一种限制性片段长度多态性（RFLP）分析方法，用于鉴定和区分旧大陆的非洲-欧亚正痘病毒（OPV）：天花病毒、痘苗病毒、牛痘病毒、猴痘病毒、骆驼痘病毒、埃可病毒和塔特痘病毒。该测试使用通过PCR从病毒基因组DNA产生的扩增子，所用引物对是根据已知分类学来源的43种不同OPV菌株的细胞因子反应调节因子B（crmB）基因的序列设计的共有引物对。该引物对从所测试的115个OPV样本中的每一个都扩增出了单一的特异性产物。大小特异性扩增子可鉴定和区分含有截短crmB基因的埃可病毒和痘苗病毒株，并使其能够与其他OPV物种区分开来。扩增产物的限制性酶切允许鉴定和区分天花病毒、猴痘病毒、骆驼痘病毒、痘苗病毒和牛痘病毒的物种和毒株。
