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利用聚合酶链反应检测禽白血病病毒J亚群

Detection of avian leukosis virus subgroup J using the polymerase chain reaction.

作者信息

Smith E J, Williams S M, Fadly A M

机构信息

U.S. Department of Agriculture, Agricultural Research Service, Avian Disease and Oncology Laboratory, Lansing, MI 48823, USA.

出版信息

Avian Dis. 1998 Apr-Jun;42(2):375-80.

PMID:9645329
Abstract

A polymerase chain reaction (PCR) assay was developed for the detection of avian leukosis virus strain J (ALV-J) in chickens. Primers were based in the E element and the 3' terminus of the long terminal repeat of proviral ALV-J. PCR products were amplified from genomic DNA extracted from chicken embryo fibroblasts (CEF) infected with either strain HPRS-103, the prototype of ALV-J, or field isolates of ALV-J obtained from broiler breeder flocks in the United States that exhibited myeloid leukosis. The newly developed PCR detected ALV-J in DNA prepared from CEF inoculated with ALV-J but not from CEF inoculated with subgroup A, B, C, D, or E. The PCR also detected ALV-J in DNA prepared from blood, combs, and toes obtained from chickens experimentally infected with ALV-J and in DNA obtained from peripheral blood monocytes from naturally infected broiler breeder chickens. The PCR described here offers a specific and sensitive alternative to conventional virus isolation tests for ALV-J.

摘要

已开发出一种聚合酶链反应(PCR)检测方法,用于检测鸡体内的禽白血病病毒J株(ALV-J)。引物基于前病毒ALV-J的E元件和长末端重复序列的3'末端。从感染了HPRS-103株(ALV-J的原型株)或从美国表现出髓性白血病的肉种鸡群中获得的ALV-J田间分离株的鸡胚成纤维细胞(CEF)提取的基因组DNA中扩增PCR产物。新开发的PCR方法能从接种了ALV-J的CEF制备的DNA中检测到ALV-J,但不能从接种了A、B、C、D或E亚群的CEF制备的DNA中检测到。该PCR方法还能从经实验感染ALV-J的鸡的血液、鸡冠和脚趾制备的DNA中以及从自然感染的肉种鸡的外周血单核细胞获得的DNA中检测到ALV-J。本文所述的PCR方法为ALV-J的传统病毒分离检测提供了一种特异且灵敏的替代方法。

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