title: Development and optimization of a biopreparedness protocol for extracting and detecting avian influenza virus in broiler chicken meat.
abstract: Detection of avian influenza virus (AIV) in poultry meat is hampered by the lack of an efficient analytical method able to extract and concentrate viral RNA prior to PCR. In this study we developed a method for extracting and detecting AIV from poultry meat by a previously standardized 1-step real-time reverse transcriptase PCR (RRT-PCR) assay. In addition, a new process control, represented by feline calicivirus (FCV), was included in the original protocol, to evaluate all analytical steps from sample preparation to the detection phase. The detection limit was below 1×10(-1) TCID50 of AIV per sample, and the quantification limit corresponded to 1×10(1) TCID50 of AIV per sample. Moreover, the addition of 1×10(2) TCID50/sample of FCV did not affect the quantification and detection limit of the reaction. These results show that the developed assay is suitable for detecting small amounts of AIV in poultry meat. In addition, the developed biopreparedness protocol can be applied to detect AIV in legal or illegal imported broiler chicken meat. The availability of a rapid and sensitive diagnostic method based on molecular identification of AIV in poultry meat provides an important tool in the prevention of AIV circulation.
authors: Simona Di Pasquale is a Technical Assistant; Emiliana Falcone is a Senior Researcher; Gabriele Vaccari is a Researcher; Dario De Medici is a Senior Resaercher; and Livia Di Trani is a Senior Researcher; all at Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità , Rome, Italy. Rickard Knutsson, PhD, is Director of Security Department, National Veterinary Institute (SVA), Uppsala, Sweden .
authors: Simona
authors: S
authors: Di Pasquale
authors: Emiliana
authors: E
authors: Falcone
authors: Rickard
authors: R
authors: Knutsson
authors: Gabriele
authors: G
authors: Vaccari
authors: Dario
authors: D
authors: De Medici
authors: Livia
authors: L
authors: Di Trani
mesh_terms: D000818:Animals; D002645:Chickens / Q000821:virology; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D057230:Limit of Detection; D008460:Meat / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014773:Virology / Q000379:methods*
chemical_list: D012367:RNA, Viral