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Dengue virus serotypic identification using suckling mouse and western blot technique
V Churdboonchart1, M Khemarunmanus, S Yoksan
1Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Abstract:
The suckling mouse which is used in the classical method to detect and propagate dengue viruses was evaluated in conjunction with the western blot and immunoenzymatic methods to detect the infecting strains of dengue viruses. After intracerebral inoculation of patients' sera into the suckling mice for 7 days, the mice were examined for the presence of dengue proteins, even though the mice did not have the neurological symptoms which usually serve as an indicator for the presence of dengue infection in the mouse brain. With a blind study of a set of 12 specimens, the suckling mice could detect the virus with the same frequency as the mosquito system but in shorter time of incubation period. The whole process to identify the type of infection takes 9 days. Another important finding is the demonstration of the virion antigen in the liver. The quantity and quality of viral proteins in liver are comparable to those in the brain suggesting that the virus may replicate in the liver as well as in the brain.
Insights
Suckling mice effectively detect dengue virus strains, even without neurological symptoms. This method identifies viral antigens in both brain and liver, suggesting dual replication sites for dengue virus.
Area of Science:
- Virology
- Immunology
- Infectious Diseases
Background:
- The suckling mouse model is a classical method for dengue virus detection and propagation.
- Traditional detection relies on observing neurological symptoms in mice, which may not always occur.
- Alternative methods like western blot and immunoenzymatic assays are used for dengue virus identification.
Purpose of the Study:
- To evaluate the efficacy of the suckling mouse model in detecting dengue virus strains.
- To compare the suckling mouse method with existing diagnostic techniques.
- To investigate dengue virus replication sites within the infected mouse.
Main Methods:
- Intracerebral inoculation of patients' sera into suckling mice.
- Incubation period of 7 days.
- Detection of dengue viral proteins using western blot and immunoenzymatic methods, independent of neurological symptoms.
- Blind study involving 12 patient specimens.
Main Results:
- Suckling mice detected dengue virus with similar frequency to mosquito systems but in a shorter incubation period.
- The entire diagnostic process, including strain identification, was completed in 9 days.
- Dengue viral antigens were detected in the liver, with protein quantity and quality comparable to the brain.
Conclusions:
- The suckling mouse model is a viable and efficient method for detecting dengue virus strains, even in the absence of overt neurological signs.
- The study demonstrates the potential for dengue virus replication in both the brain and the liver of infected suckling mice.
- This model offers a faster alternative for dengue virus diagnosis compared to traditional methods.