Use of Restriction Fragment Length Polymorphism, Immunoblotting, and Polymerase Chain Reaction in the Differentiation of Avian Poxviruses

Abstract

Abstract. Restriction deoxyribonucleic acid (DNA) fragment profile analysis coupled with immunogenic protein profile analysis has provided useful information in determining the differences between vaccine strains and field isolates of fowlpox virus (FPV). The DNA of strains examined in this study clearly fell into 3 minor groups of restriction patterns similar but distinct from one another: restriction patterns exhibited by the vaccine strains except 1 vaccine strain, Vac-82; restriction profiles indicated by Vac-82 and field isolates FI-38 and FI-42; and restriction patterns indicated by field isolates FI-43, FI-51, FI-54, and FI-56. Furthermore, when the strains were analyzed and compared by immunoblotting analysis, they showed group differences similar to the differences in restriction profiles. Both techniques provided high sensitivity in verifying differences between vaccine strains and field isolates of FPV. The disparity found in restriction fragments or immunogenic protein profile between vaccine strains and field isolates does not exclude the appreciable high degree of DNA sequence conservation and homology. However, the minor disparity observed in these strains suggests a molecular basis for why vaccinated commercial flocks could have continually been infected by variant strains of FPV. A rapid and sensitive polymerase chain reaction method, which amplified a product from the 4b core protein gene of the FPV genome, was developed for identification and differentiation of members of the genus Avipoxvirus. Whereas total DNA from either vaccine strains or field isolates was used as template for amplifying a predicted product of 578 or 1409 bp, only cleavage of the amplified product (1409 bp) represented an additional detection technique for species differentiation. An attempt to distinguish between strains on the basis of amplification product was partially successful.

Retracted:MHC diversity and differential exposure to pathogens in kestrels (Aves:Falconidae)

“…(2005 ). The presence of poxvirus (Tadese & Reed 2003), the paramyxovirus causing the Newcastle disease (Farkas et al. 2007), the serotypes H5, H7 and H9 of the avian influenza (Kiss et al.…”

Livestock Drugs and Disease: The Fatal Combination behind Breeding Failure in Endangered Bearded Vultures

“…in blood was determined as described previously [29,30]. The presence of poxvirus, the paramyxovirus causing Newcastle disease, the serotypes H5, H7 and H9 of avian influenza, falcon adenovirus, circovirus, herpesvirus, polyomavirus, reovirus and West Nile virus were determined following the PCR-based methods available in the literature [31][32][33][34][35][36][37][38][39]. We also searched for helminths and protozoans in the gastrointestinal tract by macroscopic and microscopic observations using standard protocols [40].…”

Development of a Multiplex Polymerase Chain Reaction for Differential Diagnosis of Canary Pox Virus

“…To date, most of the identification methods, concerning the APVs are based on PCR and restriction enzyme analysis of 4b gene of pox viruses, a highly conserved in all of Chordopoxvirinae. Differentiation of some APVs has been carried out by using EcoRV and NlaII restriction enzyme profiles on the amplified 4b gene fragment (4). In other study Luschow et al (14) have revealed that restriction digestion of a region between the gene with either MseI or EcoRV enzymes resulted in differentiation of APVs that showed a nucleotide similarity of 72-100%.…”

“…Variable restriction enzyme profiles suggest significant genomic differences among avipox viruses (3)(4)(5). Canarypox virus (CPV) is an etiologic agent of canarypox which produces clinical signs including both cutaneous and diptheretic disease forms caused by the prototypical FPV and includes proliferative and necrotic changes in epithelial tissues of the dermis, notably around the eyes and commissures of the beak, feet, and respiratory tract (6)(7)(8).…”