Detection of horses infected naturally with equine infectious anemia virus by nested polymerase chain reaction
| Autor: | Malliga M. Nagarajan, Carole Simard |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Immunodiffusion
Sequence analysis viruses Molecular Sequence Data Restriction Mapping Polymerase Chain Reaction Virus law.invention Equine infectious anemia law Virology Animals Humans Horses Polymerase chain reaction biology Base Sequence Sepharose Sequence Analysis DNA Group-specific antigen Provirus biology.organism_classification Molecular biology Equine Infectious Anemia Lentivirus DNA Viral Nested polymerase chain reaction Gels Infectious Anemia Virus Equine |
| Zdroj: | Journal of virological methods. 94(1-2) |
| ISSN: | 0166-0934 |
| Popis: | A nested polymerase chain reaction (PCR) amplifying a region of the gag gene of equine infectious anemia virus (EIAV) was developed for the rapid and direct detection of proviral DNA from the peripheral blood of naturally infected horses and was compared with the Coggins test. DNA prepared from white blood cells of 122 field horses from 15 stables with reported cases of EIAV and one seronegative stable were analysed. Amplifications of expected size fragments were obtained by nested PCR for 88 horses using two different sets of primers targeting the gag region. The specificity of the amplified products was confirmed by hybridization using a digoxigenin-labeled probe. Gag-nested PCR-restriction fragment length polymorphism analysis distinguished two different subtypes of gag gene, A and B. Subtype A was found to be the most prevalent among the infected horses that were tested. The PCR-gag amplified sequence of subtype A shared 84.6% nucleotide and 93% deduced amino acid sequence identities with the prototype Wyoming strain whereas subtype B sequence was almost 100% identical to the prototype. Sequence analysis of gag subtype A suggests the presence of a novel EIAV variant among infected horses in Canada. The nested PCR assay developed in the present study detected more EIAV positive animals and was found as specific as the agar gel immunodiffusion (Coggins) assay and offers great potential a diagnostic test for the detection of EIAV infections in field horses. |
| Databáze: | OpenAIRE |
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