High-performance Detection Of Hantaan Virus NP Based On Protein Chips And Immuno-PCR

Hemorrhagic fever with renal syndrome (HFRS) is zoonosis which occur worldwide, has high mortality rate and is a significant public health problem. The causative agents of HFRS are hantaviruses belonging to the Bunyaviridae family. Clinical syndrome of HFRS is complicate and quickly changeable, which brings some difficulty to clinical diagnosis. At present, serological diagnosis remain major methods, such as immunofluorescence antibody assay (IFA), but sensitivity and specificity of these methods are not satisfactory. Therefore, it is necessary to develop high-sensitive, high-specific and easy-manipulate methods to detect HFRS as assistant techniques. According to the requirements above, we develop several immunoassay methods: mainly scFv-mediated immunofluorescence protein chips and immuno-PCR. By fusing anti-NP scFv with EAP, we obtain detecting antibody making EAP as a signal molecule. Meanwhile, Cy3 labeled scFv and scFv-SBP fusion protein are utilized to detect hantavirus NP based on protein chip. Detection sensitivity of protein chips combined with fluorescence nanoparticles is 10 pg/ml and immino-PCR is 10 fg/ml, which are higher than conventional ELISA (10 μg/ml). In addition, we try rapid and simple homogenous immunoassay with a water-soluble fluorescent polymer, which can detect 10 ng/ml hantaan virus NP in several minutes.