Detection of Avian leukosis virus genome by a nested polymerase chain reaction using DNA and RNA from dried feather shafts
Detection of Avian leukosis virus genome by a nested polymerase chain reaction using DNA and RNA from dried feather shafts
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DOI：
10.1177/104063870902100415
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发表时间：
2009-07-01
影响因子：
1.5
通讯作者：
Umemura, Takashi
中科院分区：
文献类型：
--
作者：
Hatai, Hitoshi;Ochiai, Kenji;Umemura, Takashi
利用冷冻羽毛果肉的套式聚合酶链式反应(NPCR)检测禽胶质瘤诱导病毒(FGV)，该病毒属于禽白血病病毒家族，最近有研究表明FGV已传播到日本动物园饲养的观赏鸡。为了建立一种简便的组织保存方法，本研究探讨了用干羽毛干标本中的DNA和RNA作为聚合酶链式反应样品的可行性。采集7只FGV阳性鸡的羽轴，在室温下保存30d。从这些干燥材料中提取DNA和RNA。从该RNA中提取的DNA和互补DNA分别对鸡P-肌动蛋白和口蹄疫病毒呈阳性反应。对在N动物园饲养的日本家禽进行了FGV的筛查，从57只禽类采集的羽柄中，根据干羽柄的DNA和DNA样本的PCR检测，有1份样本FGV阳性。这只阳性鸟来自A动物园，其脑部病变提示为禽类胶质瘤。结果表明，从干燥羽轴中提取的DNA和RNA可用于NPCR检测口蹄疫病毒基因组。
The nested polymerase chain reaction (nPCR) using frozen feather pulp is useful for detecting fowl glioma-inducing virus (FGV), which belongs to the Avian leukosis virus family, and it has recently been suggested that FGV has spread to ornamental chickens kept in Japanese zoological gardens. In the Current study, the practicality of using DNA and RNA from dried feather shafts as PCR samples was examined to establish a simple method for tissue preservation. Feather shafts were collected from 7 FGV-positive chickens and stored at room temperature for 30 days. DNA and RNA were extracted from these dried materials. All DNA and complementary DNA (cDNA) prepared from the RNA showed positive results for chicken P-actin and FGV, respectively. Screening for FGV was performed on Japanese fowls kept in zoological garden N. Of the feather shafts collected from 57 birds, 1 sample tested positive for FGV according to PCR of DNA and cDNA samples from the dried feather shafts. This positive bird originated from zoological garden A and had brain lesions suggestive of fowl glioma. The results suggest that DNA and RNA from dried feather shafts can be used in nPCR to detect the FGV genome.