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Title: DEVELOPMENT AND VALIDATION OF A REAL-TIME RT-PCR TEST FOR AVIAN PARAMYXOVIRUS TYPE 1 AND EXOTIC NEWCASTLE DISEASE VIRUS

Author
item Spackman, Erica
item Wise, Mark
item Suarez, David
item SENNE, D - NVSL-APHIS, AMES,IA
item PEDERSEN, J - NVSL-APHIS, AMES,IA
item King, Daniel
item Swayne, David
item Lee, Chang
item Seal, Bruce

Submitted to: American Association of Veterinary Laboratory Diagnosticians
Publication Type: Abstract Only
Publication Acceptance Date: 10/9/2003
Publication Date: 10/9/2003
Citation: Spackman, E., Wise, M., Suarez, D.L., Senne, D.A., Pedersen, J.C., King, D.J., Swayne, D.E., Lee, C.W., Seal, B.S. 2003. Development and Validation of a Real-Time RT-PCR Test for Avian Paramyxovirus Type 1 and Exotic Newcastle Disease Virus. American Association of Veterinary Laboratory Diagnosticians.

Interpretive Summary:

Technical Abstract: Real-time RT-PCR tests were developed for the detection of avian paramyxovirus type-1 (APMV-1) and for the specific detection of the exotic Newcastle disease (END) virus circulating during the 2002-2003 outbreak in California, Arizona, Nevada and Texas. The test for APMV-1 targeted the matrix gene and the assay specific for END targets the fusion gene. The specificity of each test was demonstrated by assessing the ability of each test to detect RNA from a variety of virulent and avirulent APMV-1 isolates. Both assays were also compared directly with virus isolation to determine their diagnostic sensitivity (DSn) and specificity (DSp) using 1461 clinical swab samples collected during the END outbreak. The APMV-1 assay DSn and DSp was 96.7% and 97.3% respectively. The END assay had DSn of 92.9% and DSp of 99.1%. Additionally, two RNA extraction methods (Trizol reagent and Qiagen RNeasy) were compared for each of two sample types; swab material (tracheal and cloacal) and tissue samples. A total of 539 swab samples and 389 tissue samples were tested by both RNA extraction methods. The optimal RNA extraction method depended on sample type.