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Application of the polymerase chain reaction to detect fowl adenoviruses.

机译:聚合酶链反应在检测禽腺病毒中的应用。

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摘要

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.
机译:测试了使用聚合酶链反应(PCR)检测禽腺病毒(FAdV)的可能性。对纯化的8型禽腺病毒基因组DNA(FAdV-8)的最佳反应参数进行评估和定义,然后对从感染细胞中提取的核酸应用相同条件。通过琼脂糖凝胶中PCR产物的溴化乙锭染色,检测到100皮克纯化的病毒DNA或250个受FAdV-8感染的细胞。当PCR产物与特异性标记探针杂交时,灵敏度提高到10 pg纯化病毒DNA或25个感染细胞。可以使用相同的引物和条件扩增FAdV和CELO病毒(FAdV血清型1)的几种野外分离株,但扩增子的大小要小于FAdV-8 PCR产物的大小。其他禽病毒和未感染的细胞培养物呈阴性。

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