Evaluation of Flinders Technology Associates cards for storage and molecular detection of avian metapneumoviruses.
The feasibility of using Flinders Technology Associates (FTA) cards for the molecular detection of avian metapneumovirus (aMPV) by reverse transcriptase-polymerase chain reaction (RT-PCR) was investigated. Findings showed that no virus isolation was possible from aMPV-inoculated FTA cards, confirming viral inactivation upon contact with the cards. The detection limits of aMPV from the FTA card and tracheal organ culture medium were 10(1.5) median ciliostatic doses/ml and 10(0.75) median ciliostatic doses/ml respectively. It was possible to perform molecular characterization of both subtypes A and B aMPV using inoculated FTA cards stored for up to 60 days at 4 to 6°C. Tissues of the turbinate, trachea and lung of aMPV-infected chicks sampled either by direct impression smears or by inoculation of the tissue homogenate supernatants onto the FTA cards were positive by RT-PCR. However, the latter yielded more detections. FTA cards are suitable for collecting and transporting aMPV-positive samples, providing a reliable and hazard-free source of RNA for molecular characterization.
a Institute of Infection and Global Health , University of Liverpool , Neston , UK.
Faez
F
Awad
Matthew
M
Baylis
Richard C
RC
Jones
Kannan
K
Ganapathy
D000818:Animals; D002645:Chickens / Q000821:virology*; D029121:Metapneumovirus / Q000235:genetics / Q000302:isolation & purification*; D008851:Micropore Filters; D018184:Paramyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000662:veterinary*; D038301:Virus Inactivation
D012367:RNA, Viral