1.Establishment and characterization of an infectious cDNA clone of a classical swine fever virus LOM strain.
Gil Soon PARK ; Seong In LIM ; Seung Ho HONG ; Jae Young SONG
Journal of Veterinary Science 2012;13(1):81-91
Classical swine fever virus (CSFV) causes a highly contagious disease among swine that has an important economic impact worldwide. CSFV strain LOM is an attenuated virus of low virulent strain of Miyagi isolated from Japan in 1956. Eight DNA fragments representing the genome of the CSFV strain LOM were obtained by RT-PCR. These were used to determine the complete nucleotide sequence and construct a full-length cDNA clone which was called Flc-LOM. Sequence analysis of the recombinant clone (Flc-LOM) revealed the presence of eight mutations, resulting in two amino acid substitutions, when compared to the parental sequence. RNA transcripts of both LOM and Flc-LOM were directly infectious in PK-15 cells. The rescued Flc-LOM virus grew more slowly than the parental virus, LOM, in the cells. Intramuscular immunization with Flc-LOM was safe and highly immunogenic in pigs; no clinical signs or virus transmission to sentinel animals were observed after 35 days. CSFV-specific neutralizing antibodies were detected 14 days post-infection. After challenge with the virulent CSFV strain SW03, pigs immunized with Flc-LOM were shown to be fully protected. Thus, our newly established infectious clone of CSFV, Flc-LOM, could serve as a vaccine candidate.
Animals
;
Antibodies, Viral/blood
;
Base Sequence
;
Cell Line
;
Classical Swine Fever/immunology/*virology
;
Classical swine fever virus/*genetics/immunology/pathogenicity
;
Cloning, Molecular
;
DNA, Complementary/genetics/immunology
;
Immunization/methods/standards/veterinary
;
Molecular Sequence Data
;
Neutralization Tests/veterinary
;
RNA, Viral/chemistry/genetics
;
Recombinant Proteins/immunology
;
Reverse Transcriptase Polymerase Chain Reaction/veterinary
;
Sequence Analysis, DNA
;
Specific Pathogen-Free Organisms
;
Swine
;
Virulence
2.Sample type is vital for diagnosing infection with peste des petits ruminants virus by reverse transcription PCR.
Pam Dachung LUKA ; Chrisostom AYEBAZIBWE ; David SHAMAKI ; Frank Norbert MWIINE ; Joseph ERUME
Journal of Veterinary Science 2012;13(3):323-325
Peste des petits ruminants (PPR) diagnosis from suspected samples from sheep and goats was carried out. Buffy coat, tissues, and oculo-nasal swabs were analyzed using nucleoprotein (NP3/NP4) and fusion protein (F1/F2) gene primers, respectively. Analysis of the sample types and primer set revealed that buffy coat are the best type of samples for PPR diagnosis and the use of two set of primers will increase the number of positives.
Animals
;
DNA Primers/analysis
;
Eye/virology
;
Goat Diseases/blood/*diagnosis/epidemiology/virology
;
Goats
;
Hair/virology
;
Nose/virology
;
Nucleoproteins/analysis
;
Peste-des-Petits-Ruminants/blood/*diagnosis/epidemiology/virology
;
Peste-des-petits-ruminants virus/genetics/*isolation & purification
;
Pigmentation
;
RNA, Viral/genetics/*isolation & purification
;
Reverse Transcriptase Polymerase Chain Reaction/*methods/standards/veterinary
;
Sheep
;
Sheep Diseases/blood/*diagnosis/epidemiology/virology
;
Uganda/epidemiology