scholarly journals Detection and partial genetic characterisation of novel avi- and siadenoviruses in racing and fancy pigeons (Columba livia domestica)

2016 ◽  
Vol 64 (4) ◽  
pp. 514-528 ◽  
Author(s):  
Mónika Z. Ballmann ◽  
Balázs Harrach
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Dna Polymerase ◽  
Nested Pcr ◽  
Columba Livia ◽  
Hexon Gene ◽  
Polymerase Gene ◽  
Viral Dna ◽  
Phylogeny Analysis ◽  
Pcr Targeting

Up to now, only a single adenovirus (AdV) isolate seemingly specific for pigeons, hence named pigeon AdV-1 (PiAdV-1), has been characterised at DNA sequence level. In the present work, the prevalence and diversity of AdVs occurring in domestic pigeon were examined by a survey performed on randomly collected samples using a very efficient, consensus nested PCR targeting the viral DNA polymerase gene. The newly detected viruses were characterised by sequencing and phylogeny analysis. Amplification of additional genome fragments was attempted by the use of several other PCR methods aiming at the hexon gene. During a 4-year survey, samples from dead or live, healthy pigeons originating from 27 lofts were examined in Hungary. Almost 50% of the samples (48 out of 97) proved to be positive for AdV. Sequence analysis revealed the presence of four hitherto unknown pigeon AdV types. PiAdV-1 was also identified in one sample. Two novel viruses named PiAdV-2 and -3 were found to belong to the genus Aviadenovirus, and two other novel types (PiAdV-4 and -5) to the genus Siadenovirus. This is the first report on the occurrence of siadenoviruses in birds belonging to the order Columbiformes. Approximately two-thirds of the PiAdV-2 genome was sequenced and analysed.

scholarly journals Isolation, Identification and Characterization of a Novel Megalocytivirus from Cultured Tilapia (Oreochromis spp.) from Southern California, USA

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3524
Author(s):  
Khalid Shahin ◽  
Kuttichantran Subramaniam ◽  
Alvin C. Camus ◽  
Zeinab Yazdi ◽  
Susan Yun ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Southern California ◽  
Phylogenetic Analyses ◽  
Head Kidney ◽  
Cell Culture Supernatant ◽  
Post Inoculation ◽  
Polymerase Gene ◽  
European Chub ◽  
Dna Polymerase Gene ◽  
Pcr Targeting

In spring 2019, diseased four-month-old tilapia (Oreochromis spp.) from an aquaculture farm in Southern California, USA were received for diagnostic evaluation with signs of lethargy, anorexia, abnormal swimming, and low-level mortalities. At necropsy, non-specific external lesions were noted including fin erosion, cutaneous melanosis, gill pallor, and coelomic distension. Internal changes included ascites, hepatomegaly, renomegaly, splenomegaly, and multifocal yellow-white nodules in the spleen and kidney. Cultures of spleen and kidney produced bacterial colonies identified as Francisella orientalis. Homogenized samples of gill, brain, liver, spleen, and kidney inoculated onto Mozambique tilapia brain cells (OmB) developed cytopathic effects, characterized by rounding of cells and detaching from the monolayer 6–10 days post-inoculation at 25 °C. Transmission electron microscopy revealed 115.4 ± 5.8 nm icosahedral virions with dense central cores in the cytoplasm of OmB cells. A consensus PCR, targeting the DNA polymerase gene of large double-stranded DNA viruses, performed on cell culture supernatant yielded a sequence consistent with an iridovirus. Phylogenetic analyses based on the concatenated full length major capsid protein and DNA polymerase gene sequences supported the tilapia virus as a novel species within the genus Megalocytivirus, most closely related to scale drop disease virus and European chub iridovirus. An intracoelomic injection challenge in Nile tilapia (O. niloticus) fingerlings resulted in 39% mortality after 16 days. Histopathology revealed necrosis of head kidney and splenic hematopoietic tissues.

Penciclovir-Resistance Mutations in the Herpes Simplex Virus DNA Polymerase Gene

1995 ◽  
Vol 6 (5) ◽  
pp. 281-288 ◽  
Author(s):  
Henry C. Chiou ◽  
Keiko Kumura ◽  
André Hu ◽  
Kelvin M. Kerns ◽  
Donald M. Coen
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Polymerase ◽  
Active Form ◽  
Resistance Mutation ◽  
Polymerase Gene ◽  
Resistance Mutations ◽  
Viral Dna ◽  
Dna Polymerase Gene ◽  
Simplex Virus

Penciclovir is the active form of the orally available prodrug famciclovir, which is entering clinical use for herpesvirus infections. Like aciclovir, penciclovir is an acyclic guanosine analogue that is phosphorylated by viral thymidine kinase and whose triphosphate can inhibit viral DNA polymerase. We tested several well-characterized herpes simplex virus mutants with aciclovir-resistance mutations in the viral DNA polymerase gene for altered sensitivity to penciclovir. The mutants varied in their susceptibilities to penciclovir with one exhibiting 2-fold hypersensitivity, one marginal resistance and three about 3-fold resistance. Marker rescue and DNA sequencing analyses mapped the penciclovir-resistance mutation of one mutant, AraA r7, to a single base change that alters a glycine to a cysteine at residue 841 within conserved region III of α-like DNA polymerases. The results have implications for the mechanism of selective action of penciclovir, for the potential for development of resistance in the clinic, and for the substrate recognition properties of herpes simplex virus DNA polymerase.

scholarly journals Sequencing of Saccharomyces telomeres cloned using T4 DNA polymerase reveals two domains.

1990 ◽  
Vol 10 (8) ◽  
pp. 4415-4419 ◽  
Author(s):  
S S Wang ◽  
V A Zakian
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Dna Polymerase ◽  
Distal Portion ◽  
Telomeric Dna ◽  
Base Pairs ◽  
Bal31 Nuclease

By using T4 DNA polymerase rather than S1 or Bal31 nuclease to clone yeast telomeres, very little telomeric DNA is lost. These clones were used to determine the DNA sequence of virtually the entire telomeric tract. Our results demonstrated that a slightly modified version, C2-3A(CA)1-6, of the consensus derived from sequence analysis of more-internal regions (J. Shampay, J. W. Szostak, and E. H. Blackburn, Nature [London] 310:154-157, 1984) extends to the very end of the chromosome. The sequence analysis also suggests that yeast telomeres consist of two domains: the proximal 120 to 150 base pairs, which appear to be protected from processes such as recombination, degradation, and elongation, and the distal portion of the telomere, which is more susceptible to these events.

scholarly journals Investigational approach to adenoviral conjunctivitis: comparison of three diagnostic tests using a Bayesian latent class model

2018 ◽  
Vol 12 (01) ◽  
pp. 043-051 ◽  
Author(s):  
Raja Sundaramurthy ◽  
Rahul Dhodapkar ◽  
Subashini Kaliaperumal ◽  
Belgode Narasimha Harish
Keyword(s):  
Sequence Analysis ◽  
Multiplex Pcr ◽  
Sensitivity And Specificity ◽  
Latent Class ◽  
Latent Class Model ◽  
Hypervariable Region ◽  
Human Adenovirus ◽  
Hexon Gene ◽  
Class Model ◽  
Pcr Targeting

Introduction: Highly contagious adenoviral conjunctivitis represents 15-70% of all conjunctivitis worldwide. Human adenovirus (hAdV) serotypes 3,4,7,8,19 and 37 contributes to 89% of all adenoviral conjunctivitis. Accurate and rapid diagnosis of adenoviral infections at serotype level could prevent misdiagnosis, spread of disease, unnecessary antibiotic use and increased treatment costs. Methodology: Sixty-two suspected viral conjunctivitis cases were recruited from November2013-January2015. Swabs collected from inferior palpebral conjunctiva and processed for viral culture (Hep2 cell line), immunofluorescence assay (IFA) and polymerase chain reaction (PCR) (targeting hexon gene). Serotype 3,4,7,8,19 and 37 identification was carried out with an optimized multiplex-PCR (based on hypervariable region of hexon gene) and confirmed by sequence analysis. Bayesian Latent Class Model (LCM) analysis was used to compare sensitivity and specificity of three tests. Results: Adenovirus was detected in 54.8% (34/62) of cases by combination of all three methods. Culture was positive in 23/34 cases (67.6%). PCR and IFA detected adenovirus in 24 (70.5%) and 21 (61.7%) cases respectively. LCM analysis revealed, sensitivity and specificity of PCR, Culture and IFA was 77.8% and 92.4%; 72.2% and 90.8%; 67.6% and 92.9% respectively. Serotyping by multiplex-PCR showed, two cases each were hAdV3 and hAdV4, 18 hAdV8 and two remained unidentified. Results of Multiplex-PCR and sequence analysis showed 100% concordance Conclusion: LCM analysis revealed, PCR is the most appropriate method for identification. Multiplex-PCR is a simple and rapid method (serotypes identification within two days); owing its short turnaround time and accuracy, it can be used as a diagnostic tool for surveillance of adenoviral keratoconjunctivitis.

Molecular Characterization and Pathogenicity of an Indian Isolate of Duck Enteritis Virus Recovered From a Natural Outbreak

2020 ◽  
Author(s):  
Sivasankar Panickan ◽  
Satyabrata Dandapat ◽  
Jyoti Kumar ◽  
Mahesh Mahendran ◽  
Sukdeb Nandi ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Pcr Amplification ◽  
Duck Enteritis Virus ◽  
Polymerase Gene ◽  
Viral Dna ◽  
Its Sequence Analysis ◽  
Dna Polymerase Gene ◽  
Enteritis Virus ◽  
Infected Duck ◽  
Natural Outbreak

Background: Duck plague is a highly contagious viral disease reported in our country very often with significant economic loss. There are some bottlenecks with the currently used ‘Holland strain’ vaccine that involves cumbersome process of vaccine production in embryonated chicken eggs. With the future goal of development of an indigenous cell culture vaccine for duck plague, the present study is aimed at isolation of an Indian strain of DEV from a natural outbreak and its characterization for the seed virus purpose. Methods: Liver samples were collected from the suspected ducks died during a natural outbreak in Kerala and subjected to polymerase chain reaction (PCR) to confirm presence of viral DNA. The duck enteritis virus (DEV) was isolated by inoculation of PCR positive samples in embryonated duck eggs/ducklings and its pathogenicity was studied. Further, the DEV recovered from the infected duck embryo and duckling liver was confirmed by PCR amplification of the viral DNA polymerase gene and its sequence analysis. Result: Out of 12 liver samples tested eight (8) were found to be positive for duck plague by PCR. The DEV infected duck embryos and ducklings died showing typical signs and characteristic gross and microscopic lesions. PCR amplification of viral DNA targeting the DNA polymerase gene yielded amplicon of expected size of 446bp. The amplicon sequence showed 99-100% homology with other DEV isolates, thus confirming the new isolate as DEV, named as DEV/India/IVRI-2016 and the gene sequence has NCBI acc. no. KX511893.

scholarly journals Development of a PCR system for porcine cytomegalovirus detection and determination of the putative partial sequence of its DNA polymerase gene

1999 ◽  
Vol 123 (1) ◽  
pp. 177-180 ◽  
Author(s):  
B. F. WIDEN ◽  
J. P. LOWINGS ◽  
S. BELAK ◽  
M. BANKS
Keyword(s):  
Dna Polymerase ◽  
Nested Pcr ◽  
Herpes Virus ◽  
Pcr Amplification ◽  
Pcr Primers ◽  
Partial Sequence ◽  
Polymerase Gene ◽  
Porcine Cytomegalovirus ◽  
Dna Polymerase Gene ◽  
Herpès Virus

After PCR amplification with conservative cytomegalovirus primers, a 520 nucleotide putative partial sequence of the DNA polymerase gene of porcine cytomegalovirus (PCMV) was determined. Sequence comparison revealed homology to DNA polymerase genes from various beta herpes viruses and a dendrogram was constructed depicting the relationship of PCMV to other members of the Herpesviridae family. The dendrogram indicates that PCMV is indeed a beta herpes virus that is more closely related to human herpes virus types 6 and 7 than to type 5.To address the difficulties encountered during conventional PCMV detection and characterization a set of nested PCR primers were constructed which generated DNA fragments of 415 and 257 bp from the DNA polymerase gene. The nested PCR system proved specific for PCMV and provided a novel means for the detection of this poorly characterized herpes virus in pig populations, vaccines and in organs used in xenotransplantation.

Sign in / Sign up

Export Citation Format

Share Document