Tosh C, Hemadri D, Sanyal A, Pattnaik B, Venkataramanan R
Central Foot-and-Mouth Disease Laboratory, Indian Veterinary Research Institute, Mukteswar-Kumaon, Nainital, India.
Acta Virol. 1997 Jun;41(3):153-5.
A method of reverse transcription (RT) and polymerase chain reaction (PCR) amplification of 1D (VP1) gene of foot-and-mouth disease (FMD) virus using one reaction mixture containing both avian myeloblastosis virus (AMV) reverse transcriptase (RTase) and Tfl DNA polymerase is described. The procedure was time saving, made use of a single buffer for both RT and subsequent amplification and performed better than the two-step procedure usually conducted with Moloney murine leukemia virus (MMLV) RTase and Taq DNA polymerase for amplification of the VP1 gene of field isolates of FMD virus serotypes O,-A, C and Asia 1. The failure to amplify the VP1 gene of many type O and Asia 1 viruses using MMLV RTase-Taq polymerase enzyme system could be overcome by performing RT of the viral genome at a higher temperature (48 degrees C) with AMV RTase which is not possible with MMLV RTase.
描述了一种使用含有禽成髓细胞瘤病毒（AMV）逆转录酶（RTase）和Tfl DNA聚合酶的单一反应混合物进行口蹄疫（FMD）病毒1D（VP1）基因逆转录（RT）和聚合酶链反应（PCR）扩增的方法。该方法节省时间，RT和后续扩增均使用单一缓冲液，并且比通常用莫洛尼鼠白血病病毒（MMLV）RTase和Taq DNA聚合酶进行的两步法在扩增口蹄疫病毒O、A、C和亚洲1型血清型的田间分离株的VP1基因时表现更好。使用MMLV RTase-Taq聚合酶酶系统未能扩增许多O型和亚洲1型病毒的VP1基因，这可以通过用AMV RTase在较高温度（48摄氏度）下对病毒基因组进行RT来克服，而这用MMLV RTase是不可能的。
