Pennsylvania Animal Diagnostic Laboratory System
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A new procedure for avian reovirus diagnosis using cell cultures and FA staining

Huaguang Lu and Tracy Ward

Avian reovirus has been isolated frequently from clinical specimens of chickens, turkeys and Muscovy ducks submitted at ADL. A new diagnostic procedure has been developed and effectively used for isolation and identification of avian reovirus. This procedure includes 3 major steps:

  1. inoculation of clinical specimens into chicken embryo kidney/fibroblasts (CEK/CEF) cell cultures for chicken reovirus isolation, and Muscovy duck embryo kidney/fibroblasts (CEK/CEF) cell cultures for Muscovy duck reovirus isolation, all cell cultures are prepared using 25cm2 cell cultural flasks;
  2. harvest the infected cells when cytopathic effects (CPE) are obsewrved and the cells are released from the monolayer to the cultural medium;
  3. preparation of the harvested cells on glass slides, air drying, acetone fixation and FA staining.

Since only floating cells are harvested and prepared for FA staining, a diagnostic result is made without termination of cell cultures, which is important when the cell cultures are continued for screening other viruses. Furthermore, the harvested cells can be used for detection of other avian viruses, which will be discussed along with application of this procedure to other cell cultures for other virus detections.

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