University of Khartoum

Serogrouping And Topotyping Of Sudanese And United States Strains Of Epizootic Hemorrhagic Disease Virus Using Pcr

Serogrouping And Topotyping Of Sudanese And United States Strains Of Epizootic Hemorrhagic Disease Virus Using Pcr

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Title: Serogrouping And Topotyping Of Sudanese And United States Strains Of Epizootic Hemorrhagic Disease Virus Using Pcr
Author: Mukhtar, Moawia M.; Ghalib, Hashim W.
Abstract: The potential use of the recently reported polymerase chain reaction (PCR) protocol for detection of United States epizootic hemorrhagic disease virus (EHDV) serotype 1 (EHDV-1) and serotype 2 (EHDV-2) ribonucleic acid in cell culture and clinical specimens was evaluated for detection of Sudanese EHDV strains. EHDV serotype 5 (EHDV-5) and EHDV, isolate 318 (untyped) designated (EHDV-318), recovered from sentinel calves at the Khartoum University farm (Sudan) were studied. RNA from EHDV-5 and EHDV-318 and a number of EHDV field isolates, propagated in cell cultures, were detected by the described PCR-based assay. The specific 387 by PCR products were visualized on ethidium–bromide stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with non-radiolabeled internal probe. Amplification product was not detected when the PCR-based assay was applied to RNA from bluetongue virus (BTV) prototypes serotypes 2, 10, 11, 13, 16 and 17; total nucleic acid extracts from uninfected BHK-21 cells. The results of this study indicated that the previously described EHDV–PCR assay could be applied for detection of Sudanese as well as United States strains of EHDV serogroup. In addition, the described EHDV–PCR assay could be used as a supportive diagnostic assay to the current conventional virus isolation procedures used for detection of EHDV infection in susceptible ruminants. C 1997 Elsevier Science Ltd Key words: EHDV, PCR, serogroup, topotype, Sudanese. Resume—L'utilite d'un protocole base sur la reaction en chaine de la polymerase (RCP), mis au point pour la detection de l'ARN de souches americaines (serotypes 1 et 2) du virus de la maladie hemorrhagique epizootique (VMHE), fut evaluee pour la detection de souches soudanaises du VMHE. L'etude porta sur le VMHE serotype 5 (VMHE-5) et le VMHE isolat 318 (non serotype), design& VMHE-318, qui furent isoles de veaux sentinelles de is ferme de l'Universite de Khartoum (Soudan). Le protocole permit la detection de l'A RN du VMHE-5, du VMHE-318 et d'autres isolats du VMHE, propages en culture cellulaire. Les produits d'acides nucleiques obtenus par RCP, de 387 bp, furent mis en evidence sur gel agar, colore au bromure d'ethidium. La specificite de ces produits fut confirmee par hybridation chimioluminescente, utilisant une sonde non radio-marquee. Ces produits ne furent pas detectes lorsque la technique RCP fut appliquee a l'ARN des serotypes 2, 10, 11, 13, 16 et 17 du virus du Blue Tongue, ou a des extraits d'acides nucleiques de cellules BHK non-infectees. Ces resultats indiquent que la technique RCP peut detecter les souches soudanaises du VMHE, en plus des souches americaines. De plus, cette technique pourrait avoir une utilite diagnostique, complementant les methodes conventionnelles d'isolation virale du VMHE chez les ruminants. C 1997 Elsevier Science Ltd
URI: http://khartoumspace.uofk.edu/123456789/17279
Date: 2015-11-25


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