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Standardization of immunocytochemical Method for the diagnosis of Dengue Viral Infection in Aedes Aegypti Linn Mosquitoes (Diptere: Culicidae)

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Aedes aegypti is the important is the important vector of dengue heamorrhagic fever (DHF), and there are several methods for virus detection in the disadvantages of being labor-intensive and requiring fluorescent microscope as well as cryofreezer. Newer methods involving enzyme conjugates such as peroxiase in conjunction with either polyclonal or monoclonal antibodies are greatly improved. With new methods of immune-cytochemistry, it is now possible to detect dengue viral antigen in a variety of tissues. The study was aimed to standardize an immunocytochemical atreptavidin-biotin –peroxidase-complex assay for diagnosis of dengue intection in Aedes aegypti using monoclonal antibody DESC7. The infected mosquitoes were held in small cylindrical cages covered with mosquito netting,and incubated at 27 ± 1⁰C and at relative humidity of 88 ± 6%. The specificity of the immunocytochemical procedure was validated by negative and positive controls showing that antibody was bound to an approcedure was validate by negative and positive controls showing that the antibody was bound to an appropriate structure. The sensitivity and specificity were also evaluated based on Hermann’s Formula . The presence of dengue antigen on head squash preparation was detected based on ISBPC assay using monoclonal antibody against dengue. The validity and reliability of the measurement were evaluated based on kappa values, according to Landis and Koch. Positive result was detected as discrete brownish granular deposits throughout most visual fields of brain tissue. Dengue viral antigen was immunolocalized to the cytoplasm of brain cells. The immune-cytochemical test under microscope at magnification of 400x was 86.67% sensitive, 96.00% specific, and the kappa value is 0.64. Meanwhile the kappa value between two observers was 0.92, with sensitivity and specificity of 96% and 97% respectively at magnification of 1000x. The monoclonal antibody DSSC7 was sensitive, specific, valid, and reliable as primary antibody to detect dengue viral infection in Ae. Aegypti head squash preparation based on imunocytochemical streptovidin-biotin-peroxidase-complex assay under light microscope.

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Detail Information

Series Title

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Call Number

Berkala Ilmu Kedokteran, 41(1): 1--10

Publisher

Yogyakarta : FK. Universitas Dadjah Mada., 2009

Collation

10p.

Language

English

ISBN/ISSN

0126-1312

Classification

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Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

aedes
DENGUE VIRUS
CULICIDAE
IMMUNOHISTOCHEMICAL

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