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Detection of porcine circovirus type 1 in commercial porcine vaccines by loop-mediated isothermal amplification

机译:通过环介导的等温扩增检测商业猪疫苗中的猪圆环病毒1型

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摘要

A loop-mediated isothermal amplification (LAMP) method with a real-time monitoring system was developed for the detection of porcine circovirus type 1 (PCV1) in commercial swine vaccines. This method was highly specific for PCV1. No cross-reaction to porcine circovirus type 2, porcine parvovirus, pseudorabies virus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus was observed. The analytical sensitivity of the LAMP for PCV1 DNA was 10 copies/μl in the case of positive recombinant plasmid comparable to that obtained from the nested polymerase chain reaction (nested PCR). Furthermore, 25 commercial swine vaccines were tested by both the LAMP and the nested PCR, and three of them were tested positive for PCV1 DNA. These results indicate that PCV1 DNA can be real-time detected by the LAMP; the method was highly specific, sensitive, and rapid for the detection of PCV1 DNA, particularly in commercial swine vaccines.
机译:开发了一种带有实时监测系统的环介导的等温扩增(LAMP)方法,用于检测商业猪疫苗中的猪圆环病毒1型(PCV1)。该方法对PCV1具有高度的特异性。没有观察到与2型猪圆环病毒,猪细小病毒,假狂犬病病毒,经典猪瘟病毒以及猪繁殖与呼吸综合征病毒的交叉反应。在阳性重组质粒的情况下,LAMP对PCV1 DNA的分析灵敏度为10拷贝/μl,可与巢式聚合酶链反应(巢式PCR)获得的灵敏度相当。此外,通过LAMP和巢式PCR都对25种商业猪疫苗进行了测试,其中3种对PCV1 DNA呈阳性。这些结果表明,LAMP可以实时检测PCV1 DNA。该方法对PCV1 DNA的检测具有高度特异性,灵敏性和快速性,特别是在商业猪疫苗中。

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