Highly adaptable and sensitive FRET-based aptamer assay for the detection of Salmonella paratyphi A
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
Authors: Renuka, R. M.; Maroli, Nikhil; Achuth, J.; Ponmalai, Kolandaivel; Kadirvelu, K.
Abstract
Here we demonstrate a facile and versatile fluorescence resonance energy transfer (FRET) based aptasensor for rapid detection of Salmonella paratyphi A. The assay shows a detection limit up to 10 cfu.mL(-1) with no cross-reactivity with other bacterial species. Less than 8% of inter-assay coefficient variance and recovery rate between 85 and 102% attests the assay reliability. The advantages of FRET-based aptamer assay over the conventional immunoassay formats such as ELISA are the specificity, speed, reliability, and simplicity of the assay. The ssDNA aptamers specific towards pathogenic Salmonella paratyphi A were generated via whole-cell SELEX. The aptamer was conjugated onto quantum dot (QD) that served as the molecular beacon and graphene oxide (GO) was used as a fluorescence quencher. Thus the proposed method enables detection of target pathogen using FRET-based assay. Further interaction of aptamer with pathogen protein DNA gyrase was explored using classical molecular dynamics simulation. (C) 2020 Published by Elsevier B.V.
Lactobacillus plantarum surface-displayed influenza antigens (NP-M2) with FliC flagellin stimulate generally protective immune responses against H9N2 influenza subtypes in chickens
VETERINARY MICROBIOLOGY
Authors: Li, Qiong-Yan; Xu, Man-Man; Dong, Hang; Zhao, Jin-Hui; Xing, Jun-Hong; Wang, Guan; Yao, Jia-Yun; Huang, Hai-Bin; Shi, Chun-Wei; Jiang, Yan-Long; Wang, Jian-Zhong; Kang, Yuan-Huan; Ullah, Naveed; Yang, Wen-Tao; Yang, Gui-Lian; Wang, Chun-Feng
Abstract
The H9N2 avian influenza virus (AIV) causes serious economic losses to the poultry industry every year. Vaccines that induce a mucosal immune response may be successful against influenza virus infection because its transmission occurs primarily in the mucosa. To develop novel and potent oral vaccines based on Lactobacillus plantarum (L. plantarum) to control the spread of AIV in poultry industry, in the present study, we constructed and expressed fusions of the influenza antigens NP and M2 with the Salmonella Typhimurium flagellinprotein FliC on the surface of L. plantarum. Oral immunization of chicks was performed, and serum antibodies, mucosal antibodies, and specific cellular immunity were detected. Immunizing chicks with avian influenza virus was evaluated. The results showed high levels of IgG in addition to high levels of secretory immunoglobulin A (sIgA) in chickens orally administered recombinant L. plantarum. In addition, the fusion may significantly increase the levels of NP- and M2-specific T cell-mediated immunity in the case of mucosal administration of NC8-pSIP409pgsA'-NP-M2-FliC. Recombinant NC8-pSIP409-pgsA'-NP-M2-FliC mediated effectively protected chickens against influenza virus and reduced virus titers in the lung. Our study outcomes indicate that the expression of influenza NP-M2 and a mucosal adjuvant (FliC), by L. plantarum could generate a mucosal vaccine candidate for animals in the future to defend against AIVs.