1. Simultaneous identification of 6 pathogens causing porcine reproductive failure by using multiplex ligation‐dependent probe amplification
- Author
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Weihuan Fang, Zhou Bin, Wanyu Dong, Yongchun Yang, Shao Chunyan, Jiang Sheng, Houhui Song, Yang Yang, Xiaodu Wang, Yingshan Zhou, Wei Fangfang, Jing Sun, Chen Lin, Lifei Zhang, and Song Quanjiang
- Subjects
Circovirus ,musculoskeletal diseases ,congenital, hereditary, and neonatal diseases and abnormalities ,Porcine parvovirus ,Swine ,040301 veterinary sciences ,animal diseases ,viruses ,Pseudorabies ,Sensitivity and Specificity ,Virus ,0403 veterinary science ,03 medical and health sciences ,Limit of Detection ,Animals ,Porcine respiratory and reproductive syndrome virus ,Multiplex ,Multiplex ligation-dependent probe amplification ,030304 developmental biology ,Swine Diseases ,0303 health sciences ,General Veterinary ,General Immunology and Microbiology ,biology ,virus diseases ,04 agricultural and veterinary sciences ,General Medicine ,Parvovirus, Porcine ,Fetal Blood ,biology.organism_classification ,Porcine reproductive and respiratory syndrome virus ,Herpesvirus 1, Suid ,Virology ,Porcine circovirus ,Classical Swine Fever Virus ,Virus Diseases ,Classical swine fever ,Female ,Multiplex Polymerase Chain Reaction - Abstract
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.
- Published
- 2020