A real-time PCR method for detection of porcine circovirus 2
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摘要: 猪圆环病毒2型(porcine circovirus 2, PCV2),是感染猪Sus scrofa domestica的一种单链DNA病毒。建立一种快速、灵敏的检测方法,对于PCV2感染猪的筛选和疾病预防非常重要。根据PCV2 ORF2基因保守区,设计了荧光定量聚合酶链式反应(PCR)引物,利用SYBR Green作为荧光染料建立了一种定量检测PCV2的PCR方法。结果表明:该方法具有灵敏度高、特异性强和重复性好的优点,每微升的最低检测限低至101拷贝DNA。利用该方法对34份PCV2阳性临床样本进行检测,检测符合率为100%,明显高于普通PCR方法的50.0%。因此,本研究建立的PCV2实时荧光定量PCR检测方法为该疾病的预防和控制提供一种有效的检测工具。图4表4参26Abstract: To develop an accurate and rapid detection method for disease screening and prevention with porcine circovirus type 2 (PCV2), a single-strand DNA virus that infects pigs, primers targeted to the ORF2 fragment of the PCV2 conserved region were designed. A real time polymerase chain reaction (PCR) method was developed using SYBR Green as a fluorescent dye and serial dilutions of ORF2 recombinant plasmid to construct a standard curve for an absolute quantification. Results showed that the detection limit was obtained with 101 copy DNA per microliter. This method exhibited 1 000 times higher sensitivity than conventional PCR, only specific identification of PCV2, and better repeatability with the less than 2% variation coefficient of intra- or inter-assay experiments. A total of 34 PCV2 positive clinical samples were confirmed using this real time PCR method, demonstrating 100% agreement in comparison with the conventional PCR having only 50% agreement. This real time PCR method could provide a valuable tool for PCV2 prevention and control.[Ch, 4 fig. 4 tab. 26 ref.]
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Key words:
- zoology /
- porcine /
- porcine circovirus 2 /
- real-time PCR
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链接本文:
https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.2016.02.023
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