Perozo Francisco, Villegas Pedro, Estevez Carlos, Alvarado Iván, Purvis Linda B
University of Georgia (Poultry Diagnostic and Research Center), Athens, GA 30602, USA.
Avian Pathol. 2006 Apr;35(2):93-8. doi: 10.1080/03079450600597410.
The feasibility of using Flinders Technology Associates filter papers (FTA cards) to collect allantoic fluid and chicken tissue samples for Newcastle disease virus (NDV) molecular detection was evaluated. Trizol RNA extraction and one-step reverse transcriptase-polymerase chain reaction (RT-PCR) were used. FTA cards allowed NDV identification from allantoic fluid with a titre of 10(5.8) median embryo lethal doses/ml. The inactivated virus remained stable on the cards for 15 days. NDV was detected from FTA imprints of the trachea, lung, caecal tonsil and cloacal faeces of experimentally infected birds. RT-PCR detection from FTA cards was confirmed by homologous frozen-tissue RT-PCR and virus isolation. Direct nucleotide sequence of the amplified F gene allowed prediction of NDV virulence. No virus isolation was possible from the FTA inactivated samples, indicating viral inactivation upon contact. The FTA cards are suitable for collecting and transporting NDV-positive samples, providing a reliable source of RNA for molecular characterization and a hazard-free sample.
评估了使用弗林德斯技术协会滤纸（FTA卡）收集尿囊液和鸡组织样本以进行新城疫病毒（NDV）分子检测的可行性。采用了Trizol RNA提取和一步法逆转录聚合酶链反应（RT-PCR）。FTA卡能够从滴度为10（5.8）个鸡胚半数致死量/毫升的尿囊液中鉴定出NDV。灭活病毒在卡片上保持稳定15天。从实验感染鸡的气管、肺、盲肠扁桃体和泄殖腔粪便的FTA印记中检测到了NDV。通过同源冷冻组织RT-PCR和病毒分离证实了从FTA卡进行的RT-PCR检测。扩增的F基因的直接核苷酸序列能够预测NDV的毒力。无法从FTA灭活样本中分离出病毒，表明接触后病毒被灭活。FTA卡适用于收集和运输NDV阳性样本，为分子特征分析提供可靠的RNA来源且是无危害样本。
