University of Khartoum

Comparison between Hemadsorption and Hemagglutination Assays for Detection of Myxoviruses (Newcastle disease virus and Avian Influenza virus) in Infected Cell Culture

Comparison between Hemadsorption and Hemagglutination Assays for Detection of Myxoviruses (Newcastle disease virus and Avian Influenza virus) in Infected Cell Culture

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Title: Comparison between Hemadsorption and Hemagglutination Assays for Detection of Myxoviruses (Newcastle disease virus and Avian Influenza virus) in Infected Cell Culture
Author: Albukhari, Randa Elamin Abeid
Abstract: This study was conducted to compare between the Hemadsorption (HAD) and heamagglutination assays (HA) to detect myxoviruses (Newcastle and avian influenza viruses) in cell culture. The strains of Newcastle disease viruses and avian influenza virus were supplied by virology Laboratory, Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum. All viruses were inoculated in Embryonated chicken eggs for propagation and titration. Then the viruses were inoculated into cell culture (Vero cells). Ten fold serial dilutions (10-1 -10-5) from each virus were prepared in phosphate buffered saline (PBS). Each dilution was inoculated into two wells of 24 well tissue culture plate monolayers and incubated at 37°C for 7, 16, 20, 24 and 48 hours with two wells acting as control for each incubation time. After certain incubation time for each concentration supernatant was screened with heamagglutination assay for the presence of the viruses by addition of equal volumes from removed tissue culture medium from the well under test and 0.4% washed chicken red blood cells (RBCs). All wells were rinsed with phosphate buffered saline and heamadsorption assay was held spontaneously by adding 0.5ml of 0.4% washed chicken RBCs to each well, of certain incubation time then the plates were held at 4°C for 20 minutes and examined under inverted microscope for the presence of heamadsorption pattern. Result revealed that all incubation times and concentration showed positive results for the (HAD) except for the lowest concentration 10-5 of incubation time (7hrs) for both viruses. Incubation times more than 7 hrs showed increasing in hemadsorption reaction of viruses when the concentration of virus were increased. In the other hand the heamagglutination test showed negative result for all incubation times and viral concentrations (for both viruses) until they were incubated for 48 hrs. From the result it can concluded that the heamadsortion is rapid comparing to heamagglutination in detection of myxovirus in cell culture as the myxoviruses did not show cytopathic effect (CPE) easily in cell culture.
URI: http://khartoumspace.uofk.edu/123456789/26107


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