We developed a multiplex ligation‐dependent probe amplification (MLPA) assay for the simultaneous detection of 6 clinically relevant viral pathogens causing porcine reproductive failure, that is porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV) and porcine parvovirus (PPV). The limits of detection for the assay varied among the 6 target organisms from 1 to 8 copies per MLPA assay. The MLPA assay was evaluated with 346 heparinized porcine umbilical cord blood specimens, and the results of the assay were compared to those of real‐time PCR. The MLPA assay showed specificities and sensitivities of 99.2% and 100%, respectively, for PRRSV; 100% and 100%, respectively, for CSFV, PCV2, PRV and PPV. No sample was found to be positive for JEV by either the MLPA assay or the real‐time PCR. In conclusion, the MLPA assay has comparable clinical sensitivity to that of real‐time PCR assay and provides a useful tool for fast screening porcine reproductive failure‐associated viruses.

中文翻译：

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通过多重连接依赖探针扩增同时鉴定引起猪繁殖衰竭的6种病原体。

我们开发了一种多重连接依赖探针扩增（MLPA）分析方法，用于同时检测导致猪繁殖衰竭的6种临床相关病毒病原体，即猪繁殖与呼吸综合征病毒（PRRSV），日本脑炎病毒（JEV），经典猪瘟病毒（CSFV），2型猪圆环病毒（PCV2），伪狂犬病病毒（PRV）和猪细小病毒（PPV）。该检测的检测限在6种目标生物之间变化，每个MLPA检测从1到8个拷贝。使用346例肝素化的猪脐带血标本评估MLPA测定，并将测定结果与实时PCR进行比较。MLPA分析显示对PRRSV的特异性和敏感性分别为99.2％和100％。对于CSFV，PCV2，PRV和PPV，分别为100％和100％。MLPA分析或实时PCR均未发现JEV阳性。总之，MLPA分析的临床敏感性可与实时PCR分析相媲美，并为快速筛选与猪繁殖衰竭相关的病毒提供了有用的工具。