Canine coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the causative agents of gastroenteritis in dogs. Seventy fecal samples from dogs with signs of gastroenteritis (vomiting and diarrhea), twenty-five fecal samples from healthy dogs and one CPV-2 vaccine strain were amplified by semi-nested polymerase chain reaction (PCR) and semi-nested reverse transcriptase polymerase chain reaction (RT-PCR), aimed at specifically studying the gene encoding the most abundant capsid protein VP2 of CPV-2 and spike protein of CCV. The specificity of the CCV RT-PCR product was evaluated by sequencing. Positive specimens comprised 44 samples (62.8%) and 9 samples (12.8%) for CPV-2 and CCV, respectively. In nine CCV positive samples, seven displayed co-infection between CCV and CPV-2. Our CCV sequence (AF482001) showed a 94.9% nucleotide identity to CCV reported in GenBank accession number D13096. High prevalence of CCV and CPV-2 infections was found in 1–2 month- and 3–6 month-old dogs, respectively. Molecular biology of these viruses is important primarily for epidemic control and preventive measures.
A male orangutan suffered from ulcers at the buccal mucosa. We obtained swab fluid from the base of both vesicles and ulcers and collected blood for further separation into serum, plasma and peripheral blood mononuclear cells (PBMC) for detection of antibody to herpesvirus by serology and herpesvirus DNA by polymerase chain reaction (PCR) using consensus degenerate primers. Serology was positive for human EBV IgG but negative for Epstein-Barr virus (EBV) immunoglobulin (IgM), as well as for both human cytomegalovirus and herpes simplex virus IgG and IgM. Upon PCR, we obtained a 232-bp product of virus DNA from PBMC, but not from lesions, serum or plasma. We confirmed the positive result by direct sequencing and compared the nucleotide sequence with other nucleotide sequences applying the BLAST program from GenBank. The sequence was similar to lymphocryptovirus of macaque (93%), marmoset (93%), gorilla (90%) and human EBV (90%). We aligned this sequence with other sequences in GenBank and performed phylogenetic analysis, showing that it probably belongs to the gammaherpesvirus group.
Background: Herpesviruses are not only infectious agents of worldwide distribution in humans, but have also been demonstrated in various non-human primates as well. Seventy-eight gibbons were subjected to serological tests by ELISA for herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), Epstein-Barr virus (EBV) and cytomegalovirus (CMV).
1The family Parvoviridae comprises two subfamilies: Parvovirinae, which infects vertebrates and is classified into three genera, Parvovirus, Erythrovirus and Dependovirus; and Densovirinae, which infects insects and is further classified into three genera, Densovirus, Iteravirus and Contravirus. 2,3 Parvovirus is among the smallest animal DNA viruses, with the virion exhibiting a diameter of between 18 and 26 nm. The genome is comprised of single-stranded DNA of approximately 5,000 bases encoding two structural (VP1 and VP2) and two non-structural (NS1 and NS2) proteins. ABSTRACT Canine Parvovirus (CPV), a member of the genus Autonomous Parvovirus, is a non-enveloped, single-stranded DNA virus of approximately 5 kb. The virus was first described in 1978, with the original isolate being termed CPV type 2 (CPV-2). In 1979, a variant CPV strain designated CPV type 2a (CPV-2a) started to become widespread, followed by a further antigenic variant designated CPV type 2b (CPV-2b), which emerged in 1984. In due course, CPV-2b replaced CPV-2a. The vaccines presently in use are CPV-2 strain specific. Hence, the objective of the present study was to detect and genotype CPV by polymerase chain reaction (PCR), using primer sequences derived from the conserved VP2 region of the genome, and to subsequent restriction fragment length polymorphism (RFLP) analysis of the PCR product. The RFLP analysis employed the endonucleases Rsa I and Hph I in order to differentiate the CPV-2 antigenic variants and establish their distribution in Thailand. We investigated 55 fecal samples from dogs with signs of enteritis, 55 samples from healthy dogs and CPV-2 strain genotype vaccine. Thirty-four out of the 55 specimens (61.8%) from dogs with enteritis were found to be CPV DNA positive. None of the specimens from healthy dogs provided evidence of CPV DNA. After establishing the difference between wild and vaccine strains using RFLP, we found that all virus strains in our study were either CPV-2a or CPV-2b type, which differed from the vaccine strain (CPV-2). Molecular characterization and CPV typing are crucial in epidemiological studies for future prevention and control of the disease.
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