Development of a nanoparticle-assisted PCR assay for detection of porcine epidemic diarrhea virus

• Published in: Journal of virological methods Vol. 220; pp. 18 - 20
• Main Authors: Yuan, Wanzhe, Li, Yanan, Li, Peng, Song, Qinye, Li, Limin, Sun, Jiguo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.08.2015
• Subjects: Animals; Coronavirus Infections - diagnosis; Coronavirus Infections - veterinary; Coronavirus Infections - virology; Detection; DNA Primers - genetics; Molecular Diagnostic Techniques - methods; Nanoparticles; nanoPCR; Nucleocapsid Proteins - genetics; Polymerase Chain Reaction - methods; Porcine epidemic diarrhea virus; Porcine epidemic diarrhea virus (PEDV); Porcine epidemic diarrhea virus - genetics; Porcine epidemic diarrhea virus - isolation & purification; RT-PCR; Sensitivity and Specificity; Short Communication; Swine; Swine Diseases - diagnosis; Swine Diseases - virology; Veterinary Medicine - methods; Detection; Porcine epidemic diarrhea virus (PEDV); RT-PCR; nanoPCR
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2015.04.008

•A nanoPCR method was developed for the detection of PEDV.•The nanoPCR assay was 100-fold more sensitive than a conventional RT-PCR assay.•The lower detection limit was 2.7×10−6ng/μL of PEDV RNA.•This test could be applied for clinical diagnosis and field surveillance of PEDV. Porcine epidemic diarrhea virus (PEDV) is an important pig pathogen that can cause vomiting, diarrhea, and dehydration, leading to serious damage to the swine industry worldwide. In this study, a nanoparticle-assisted polymerase chain reaction (nanoPCR) assay targeting the N gene of PEDV was developed and the sensitivity and specificity were investigated. Under the optimized conditions for detection of PEDV RNA, the nanoPCR assay was 100-fold more sensitive than a conventional RT-PCR assay. The lower detection limit of the nanoPCR assay was 2.7×10−6ng/μL of PEDV RNA and no cross-reaction was observed with other viruses. This is the first report to demonstrate the application of a nanoPCR assay for the detection of PEDV. The sensitive and specific nanoPCR assay developed in this study can be applied widely in clinical diagnosis and field surveillance of PEDV-infection.