分子生物学
GenBank公司
实时聚合酶链反应
生物
聚合酶链反应
琼脂糖凝胶电泳
塔克曼
底漆(化妆品)
聚合酶
底漆二聚体
复式(建筑)
基因组DNA
基因
检出限
DNA
遗传学
化学
色谱法
多重聚合酶链反应
水热
有机化学
作者
Maya Shofa,Akiho Ohkawa,Tamaki Okabayashi,Yasuyuki Kaneko,Akatsuki Saito
标识
DOI:10.1177/10406387231154610
摘要
Domestic cat hepadnavirus (DCH) is a novel hepadnavirus, first identified in 2018. DCH is generally detected using conventional PCR assays, which include time-consuming agarose gel electrophoresis. We developed a rapid, sensitive, and specific real-time PCR (rtPCR) assay for the detection of the DCH genome. To streamline the procedure, our rtPCR assay was carried out using blood samples, without DNA extraction. A consensus primers/probe set was designed based on the nucleotide sequences of the surface/polymerase gene of all DCH strains available in GenBank. To exclude the possibility that the PCR reaction was blocked by anticoagulants, we also used a primers/probe set for amplifying the cat beta-actin gene as a reference gene. Our direct duplex rtPCR assay had high sensitivity, with a limit of detection of 10 copies/μL of blood for DCH. Our direct duplex rtPCR assay should be a useful tool for DCH detection and surveillance.
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