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Title: PROTECT YOURSELF AND YOUR SAMPLE: PROCESSING ARBOVIRUS INFECTED BITING MIDGES FOR VIRAL DETECTION ASSAYS AND DIFFERENTIAL EXPRESSION STUDIES

Author
item Drolet, Barbara
item CAMPBELL, COREY
item MECHAM, JAMES

Submitted to: Anthology of Biosafety, VI: Arththropod-Borne Diseases
Publication Type: Book / Chapter
Publication Acceptance Date: 10/25/2002
Publication Date: 11/3/2003
Citation: Drolet, B.S., Campbell, C.L., Mecham, J.O. 2003. Protect yourself and your sample: processing arbovirus infected biting midges for viral detection assays and differential expression studies. Anthology of Biosafety, VI: Arththropod-Borne Diseases.

Interpretive Summary: Arthropod-borne viral (arboviral) diseases are maintained in nature in a cycle propagated between susceptible biting insects and hosts. A susceptible biting midge, takes a blood meal from an arbovirus-infected animal, may amplify the virus, and during a subsequent feeding may transmit the virus to a naïve vertebrate host. Considerable procedural information is available for infecting Culicoides spp. with blood-borne arboviruses by artificial blood feeders (Hunt and McKinnon, 1990) or by intrathoracic injection, as well as the physical containment of these insects (Hunt and Tabachnick, 1996). However, the specifics of proper handling, processing and storage of arboviral infected insects for virus detection assays and differential expression studies are not readily available in a single source. It is crucial that these procedures are done properly to ensure effective, specific and reproducible results. It is also critical that the laboratory worker remain safe during these procedures. This chapter discusses the handling, processing and storage methods which precede various arbovirus detection and gene expression assays, while maintaining biosafety practices.

Technical Abstract: Arthropod-borne viral (arboviral) diseases are maintained in nature in a cycle propagated between susceptible biting insects and hosts. A susceptible biting midge, takes a blood meal from an arbovirus-infected animal, may amplify the virus, and during a subsequent feeding may transmit the virus to a naïve vertebrate host. Considerable procedural information is available for infecting Culicoides spp. with blood-borne arboviruses by artificial blood feeders (Hunt and McKinnon, 1990) or by intrathoracic injection, as well as the physical containment of these insects (Hunt and Tabachnick, 1996). However, the specifics of proper handling, processing and storage of arboviral infected insects for virus detection assays and differential expression studies are not readily available in a single source. It is crucial that these procedures are done properly to ensure effective, specific and reproducible results. It is also critical that the laboratory worker remain safe during these procedures. This chapter discusses the handling, processing and storage methods which precede various arbovirus detection and gene expression assays, while maintaining biosafety practices.