Development of Immune Chromatographic Test Strip for Rapid Detection of Zika Virus in Humans

Abstract

newline x newlineAbstract newlineThe present invention relates to antigen immune-chromatographic test kit for rapid detection of Zika virus using stable gold nanoparticles and its manufacturing method. Herein, we have developed an alternate viral NS1 antigen-based rapid tests that use monoclonal antibody pairs to detect and distinguish the DENV and CHKV stereotypes, as well as ZIKV, without cross-reactivity in 5 minutes. In order to mask the non-specific results due to DENV and CHKV, we have developed the Blockade of Binding (BoB) assay in lateral flow kits. Also, for better performance of kits sample pad having a minimum absorption capacity of 20 and#956;l/cm2 and NCM having a pore size of 5 15 and#956;m were used. The NCM comprises test line (TL) coated with 1 and#956;g/L of Zika NS1 monoclonal antibody and 0.4 mg/ml immobilizing capture antibodies of Goat-anti-mouse monoclonal antibody (GAM Mab) at control line (CL). The conjugate pad was having 5 and#956;g/L of anti-Zika NS1 monoclonal antibody and anti-dengue monoclonal antibody (anti-DENV-MAb) and anti-chikengunia monoclonal antibody (anti-CHKV-MAb) blockers in 0.01 and#956;g/L concentrations. Herein, AuNP-ZIKA NS1 monoclonal antibody is capable of binding to specific sites of specimen s Zika antigen. Whereas, anti-DENV-MAb is capable of binding to specimens specific site for dengue, anti-CHKV-MAb is capable of binding to specimens specific site for chikengunia, ZIKA NS1 monoclonal antibody is capable of binding to anti-ZIKA NS1 monoclonal antibody and GAM MAb is capable of binding to leftover anti-ZIKA NS1 monoclonal antibody. This test kit also has better scale up measures. The strips process capability (Cp) and process capability index (CpK) value is gt1, specificity and sensitivity calculated for entire stability period were found 95.83 % and 100 % respectively. The strips also have a 100 % positive predictive value (uniformity) for the shelf life of 12 months. newlineKeywords: ZIKV antigen test strips, Point-Of- Care, Lateral Flow, ZIKA NS1, ZIKA Virus.