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利用克隆的 cDNA 探针检测传染性法氏囊病病毒

Detection of infectious bursal disease viruses by using cloned cDNA probes.

作者信息

Jackwood D J, Kibenge F S, Mercado C C

机构信息

Food Animal Health Research Program, Ohio State University, Wooster 44691.

出版信息

J Clin Microbiol. 1989 Nov;27(11):2437-43. doi: 10.1128/jcm.27.11.2437-2443.1989.

Abstract

A molecular clone representing 445 base pairs at the 3' end of infectious bursal disease virus (IBDV) genome segment B was used in a dot blot hybridization assay to detect viral RNA from cell culture and from chicken bursa and spleen tissue specimens. The cloned nucleotide sequence represents approximately 14% of the virus-encoded polymerase (VP-1) gene. The lower detection limit of radiolabeled probes prepared from this clone was 0.1 ng of IBDV double-stranded RNA. The probe had broad specificity and was used to detect four serotype 1 IBDV strains and one serotype 2 IBDV strain. This probe, however, did not cross-react with nucleic acid extracted from nine unrelated poultry viruses. A rapid procedure for isolation of IBDV genomic RNA from bursa and spleen tissue specimens was developed and used with the dot blot hybridization assay to detect IBDV strains in tissue samples from experimentally infected and commercially reared chickens.

摘要

一个代表传染性法氏囊病病毒(IBDV)基因组B片段3'端445个碱基对的分子克隆,被用于斑点杂交试验,以检测来自细胞培养物以及鸡法氏囊和脾脏组织标本中的病毒RNA。该克隆的核苷酸序列约占病毒编码的聚合酶(VP-1)基因的14%。用此克隆制备的放射性标记探针的最低检测限为0.1 ng IBDV双链RNA。该探针具有广泛的特异性,用于检测4株1型IBDV毒株和1株2型IBDV毒株。然而,该探针与从9种不相关的家禽病毒中提取的核酸没有交叉反应。开发了一种从法氏囊和脾脏组织标本中快速分离IBDV基因组RNA的方法,并将其与斑点杂交试验一起用于检测来自实验感染鸡和商业饲养鸡的组织样本中的IBDV毒株。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8146/267053/f11cc818bd6f/jcm00071-0055-a.jpg

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