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聚合酶链反应在检测禽腺病毒中的应用。

Application of the polymerase chain reaction to detect fowl adenoviruses.

作者信息

Jiang P, Ojkic D, Tuboly T, Huber P, Nagy E

机构信息

Department of Pathobiology, Ontario Veterinary College, University of Guelph.

出版信息

Can J Vet Res. 1999 Apr;63(2):124-8.

Abstract

The possibility of using the polymerase chain reaction (PCR) for the detection of fowl adenoviruses (FAdV) was tested. The optimal reaction parameters were evaluated and defined for purified genomic DNA of type 8 fowl adenovirus (FAdV-8), and then the same conditions were applied for nucleic acid extracted from infected cells. One hundred picograms of purified viral DNA, or 250 FAdV-8-infected cells, were detected by ethidium bromide staining of the PCR products in agarose gels. The sensitivity was increased to 10 pg purified viral DNA, or 25 infected cells, when the PCR products were hybridized with a specific labeled probe. Several field isolates of FAdV and the CELO virus (FAdV serotype 1) could be amplified by the same primers and conditions, but the size of the amplicons was smaller than that for the FAdV-8 PCR product. Other avian viruses and uninfected cell cultures tested negative.

摘要

对使用聚合酶链反应(PCR)检测禽腺病毒(FAdV)的可能性进行了测试。针对8型禽腺病毒(FAdV-8)的纯化基因组DNA评估并确定了最佳反应参数,然后将相同条件应用于从感染细胞中提取的核酸。通过对琼脂糖凝胶中PCR产物进行溴化乙锭染色,可检测到100皮克纯化的病毒DNA或250个感染FAdV-8的细胞。当PCR产物与特异性标记探针杂交时,灵敏度提高到10皮克纯化的病毒DNA或25个感染细胞。几种FAdV的野外分离株和CELO病毒(FAdV血清型1)可通过相同引物和条件进行扩增,但扩增子大小比FAdV-8 PCR产物的小。测试的其他禽病毒和未感染细胞培养物均为阴性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/45c2/1189531/b427e44c8d17/cjvetres00010-0047-a.jpg

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