Anti-CSFV E2 monoclonal antibody, clone LPN24

Early diagnosis and intervention of livestock epidemics is extremely important because disastrous outbreaks of an infectious disease can not only hit livestock industry but also cause a series of negative economic and social consequences. In order to fulfill the requirement of point-of-care testing, a reliable, field-portable, time-efficient and cost-effective microfluidic platform including a microfluidic chip and a semi-automated immunoassay system (SAIS) is developed. Multiple samples can be tested simultaneously in a single chip for multiple-biomarker testing. The entire assay, including reagent injection, well washing, waste collection and colorimetric detection, can be realized in the user-friendly system. The assay of three contagious diseases, PRRSV, CSFV and PCV2, is conducted in the microfluidic platform. The consistency of the reaction for different samples can be guaranteed owing to the chip design for flow balance. The assay only takes 12 min and 4 mu L of sample is required. The assay achieves high accuracy ( > 93.8 %), high sensitivity ( > 88.8 %) and high specificity ( > 96.6 %). The platform has the potential to be applied for on-site rapid screening livestock diseases.

The non-structural 5A (NS5A) protein of classical swine fever virus (CSFV) is proven to be involved in viral replication and can also modulate cellular signaling via to its ability to interact with various cellular proteins. Here, HSP70/NS5A complex formation is confirmed by coimmunoprecipitation and GST-pulldown studies. Additionally, the N-terminal amino acids (29-240) of NS5A were identified as the interaction region through in vivo deletion analyses, and confocal microscopy showed that NS5A and HSP70 colocalized in the cytoplasm. Overexpression of HSP70 via the eukaryotic expression plasmid pDsRED N1 or lentivirus significantly promoted viral RNA synthesis. Whereas the knockdown of HSP70 by lentivirus-mediated shRNA or inhibition by quercetin markedly decreased the viral load. These data suggest that HSP70 plays a critical role in the viral life cycle, particularly during the virus RNA replication period. The investigation of HSP70 protein functions may be beneficial for developing new strategies to treat CSFV infection. (C) 2015 Elsevier Inc. All rights reserved.