Polymerase chain reaction combined with fluorescent lateral flow immunoassay based on magnetic purification for rapid detection of canine parvovirus 2

Zhuang, Linlin; Ji, Yongxin; Tian, Peilong; Wang, Kaixuan; Kou, Chengkun; Gu, Ning; Zhang, Yu

In this study, we describe the development and validation of a sensitive and specific PCR method combined with fluorescent lateral flow immunoassay (PCR-LFIA) based on magnetic beads purification for rapid detection of CPV-2 ( Fig. 1) . The LFIA used nanoparticles as signal amplifier, which ensures the sensitivity and accuracy to replace gel electrophoresis. With the LFIA reader Nanoeasy 1700, results can be quantified and shared in real time via the internet. The PCR-LFIA assay is rapid, sensitive, specific and safe, indicating great promise as a convenient tool to facilitate disease outbreak investigations and response timely. Sixty-two nasal/oropharyngeal and faecal swabs were collected from 51 dogs of both sexes (various breeds and ages) from the animal hospital of Jubilancy, Nanjing and 11 dogs from the farms in Jiangsu Province, China during 2018 and snap-frozen for storage at − 80°C. These swabs were collected from diarrheic dogs and nine of them were suspected for CPV infection. All samples were collected using sterile cotton swabs and were immersed immediately in screw capped polypropylene tubes containing sterile PBS of pH 7.2 [31] .