Nylon membrane-immobilized PCR for detection of bovine viruses
Nylon membrane-immobilized PCR for detection of bovine viruses
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DOI：
10.2144/02321st03
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发表时间：
2002-01-01
期刊：
影响因子：
2.7
通讯作者：
Melcher, U
中科院分区：
文献类型：
--
作者：
Onodera, K;d'Offay, J;Melcher, U
桥接技术（Bridge Technology， TM）是一种将引物对固定在固体载体上，只在引物对点处扩增的扩增技术。该技术具有诊断潜力，因为一系列引物对（每个引物对一种不同的病原体具有特异性）可以与诊断样本一起使用，而不存在困扰多重pcr发展的对间相互作用。因此，一个分析应该能够确定哪些多种病原体存在，哪些不存在于每个样品。作为实验材料，我们检测了含rna的牛病毒性腹泻病毒（BVDV）和两种含dna的牛疱疹病毒1和2 （BHV-1和BHV-2）的检测特异性。采用尼龙膜，分别为BVDV和bhv，用两种对应的模板进行扩增，每种模板单独扩增和无模板扩增。当化学发光检测纳入的digi -核苷酸进行扩增时，获得了预期的扩增模式。
Bridge Technology(TM) is an amplification technique in which pairs of primers are immobilized on a solid support, allowing amplification only at the location of the primer pair spot. The technique has diagnostic potential since an array of primer pairs, each specific for a different pathogen, can be used with a diagnostic sample without inter-pair interactions that plague the development of multiplex PCRs. As a result, one assay should be able to determine which of multiple pathogens are present and which are absent in each sample. As test material, we examined the specificity of detection of the RNA-containing bovine viral diarrhea virus (BVDV) and two DNA-containing bovine herpesviruses 1 and 2 (BHV-1 and BHV-2). Nylon membranes with two spots of UV-immobilized primer pairs-one for BVDV and one for BHV-were used in amplification with both corresponding templates, with each template singly and with no template. When amplification was assayed by chemiluminescent detection of incorporated DIG-nucleotides, the expected amplification patterns were obtained.