Singh Pratik, Kim Tae-Joong, Tripathy Deoki N
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802, USA.
J Vet Diagn Invest. 2003 Jan;15(1):50-4. doi: 10.1177/104063870301500111.
The use of 2 monoclonal antibodies (MAbs), P1D9 and P2D4, which recognize different fowlpox virus (FPV) antigens, for the identification and characterization of FPV strains was evaluated. Initially, the MAbs were used in conjunction with a dot blot assay that enabled FPV to be differentiated from the avian herpesvirus, infectious laryngotracheitis virus. Confirmation of the specificity of these MAbs was provided by the demonstration that only FPV antigens were recognized by a combination of both antibodies when used for immunoblotting proteins contained in various avipoxviruses. Later, an antigenic characterization of 11 FPV field isolates, 6 FPV vaccine strains, and 3 pigeonpox virus vaccines was performed by Western blotting with the individual MAbs. Whereas MAb P2D4 consistently recognized a protein with an apparent molecular weight of 60 kD, there was variability in the size of the antigen that was immunoreactive with the other MAb. For example, MAb P1D9 recognized an antigen of apparent molecular weight of 46 kD in all vaccine strains except 2 of FPV origin. In these exceptions, either only a 39-kD or both a 42- and 46-kD protein were immunoreactive. As for the field isolates, a 39-kD antigen was recognized in 8 of them, whereas a 42-kD antigen was detected in the remaining 3. Therefore, the more extensive immunoblotting technique may facilitate FPV strain differentiation, whereas routine diagnosis of fowlpox could be accomplished by using the MAb-based dot blot assay.
评估了使用2种单克隆抗体(MAb)P1D9和P2D4来鉴定和表征禽痘病毒(FPV)毒株,这两种抗体可识别不同的禽痘病毒抗原。最初,这些单克隆抗体与斑点印迹法结合使用,该方法能够将禽痘病毒与禽疱疹病毒、传染性喉气管炎病毒区分开来。当用于免疫印迹各种禽痘病毒中所含蛋白质时,只有禽痘病毒抗原能被两种抗体的组合识别,这证明了这些单克隆抗体的特异性。后来,通过用单个单克隆抗体进行蛋白质印迹法,对11株禽痘病毒野外分离株、6株禽痘病毒疫苗株和3种鸽痘病毒疫苗进行了抗原特性分析。单克隆抗体P2D4始终识别一种表观分子量为60 kD的蛋白质,而与另一种单克隆抗体发生免疫反应的抗原大小存在差异。例如,单克隆抗体P1D9在除2株禽痘病毒来源的疫苗株外的所有疫苗株中识别出一种表观分子量为46 kD的抗原。在这些例外情况中,要么只有一种39-kD的蛋白质具有免疫反应性,要么42-kD和46-kD的蛋白质都具有免疫反应性。至于野外分离株,其中8株识别出一种39-kD的抗原,而其余3株检测到一种42-kD的抗原。因此,更广泛的免疫印迹技术可能有助于禽痘病毒毒株的区分,而禽痘的常规诊断可以通过基于单克隆抗体的斑点印迹法来完成。