The SYBR Green І-based duplex real-time PCR assay was developed for simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 3 (PCV-3) genomes. PRRSV and PCV-3 were distinguished in the same sample by their distinctive melting temperature (Tm) which was 84 °C for PRRSV and 81.5 °C for PCV-3, and other non-targeted swine viruses showed no specific melting peaks. The detection limits of this assay were 46.1copies/μL for PRRSV and 49.3copies/μL for PCV-3, respectively. Thirty-three lung samples of porcine with respiratory and reproductive failure symptoms were collected and confirmed by the SYBR Green І-based real-time PCR assay and conventional PCR assay. The real-time PCR detection results showed that the PRRSV positive rate was 45.45%, the PCV-3 positive rate was 63.63%, the PRRSV and PCV-3 co-infection positive rate was 36.36%, which were more sensitive than conventional PCR detection. This duplex real-time PCR assay could be a rapid, sensitive and reliable method for the detection of PRRSV and PCV-3 co-infection.

中文翻译：

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通过基于SYBR Green的双重实时PCR同时检测猪繁殖与呼吸综合征病毒和猪圆环病毒3。

开发了基于SYBR Green的双重实时PCR分析方法，用于同时检测猪繁殖与呼吸综合征病毒（PRRSV）和猪圆环病毒3（PCV-3）基因组。PRRSV和PCV-3在同一样品中的区别在于其独特的解链温度（Tm），PRRSV为84°C，PCV-3为81.5°C，其他非靶向猪病毒均未显示特定的解链峰。该测定的检出限分别为PRRSV 46.1拷贝/μL和PCV-3 49.3拷贝/μL。收集了33例具有呼吸和生殖衰竭症状的猪肺样本，并通过基于SYBR Green的实时PCR分析和常规PCR分析进行了确认。实时PCR检测结果表明，PRRSV阳性率为45.45％，PCV-3阳性率为63.63％，PRRSV和PCV-3共感染阳性率为36.36％，比常规PCR检测更为敏感。这种双工实时PCR测定法可能是一种快速，灵敏和可靠的检测PRRSV和PCV-3合并感染的方法。