Asset Details
Do you wish to reserve the book?
Japanese encephalitis genotype I virus-like particles stably expressed in BHK-21 cells serves as potential antigen in JE IgM ELISA
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Japanese encephalitis genotype I virus-like particles stably expressed in BHK-21 cells serves as potential antigen in JE IgM ELISA
Do you wish to request the book?
Japanese encephalitis genotype I virus-like particles stably expressed in BHK-21 cells serves as potential antigen in JE IgM ELISA
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Japanese encephalitis genotype I virus-like particles stably expressed in BHK-21 cells serves as potential antigen in JE IgM ELISA
2022
Overview
Japanese encephalitis virus (JEV) is one of the leading causes of epidemic encephalitis in South Asian countries. Due to the short-term viremia, detecting IgM antibodies by ELISA is treated as the front-line diagnostic assay. Co-circulation and multiple exposures to antigenically cross-reactive flaviviruses in India pose a challenge in serodiagnosis. Replacing the whole virus antigen currently used in the JE IgM detection kits (ELISA) may improve the specificity and sensitivity of the existing JE MAC ELISA kits. For this purpose, we developed a stably transfected cell clone, BHK-IE6, which expresses a high amount of VLPs up to 37 µg/ml and is consistent in expression up to 40 passages. For the expression of VLPs in the secretory form, we cloned the JEV G-I prM-E coding gene along with the C-terminal signal sequence of capsid protein in the BHK-21 cells using the pcDNA3.1 + mammalian expression vector. The immune assays performed demonstrated its immune reactivity equivalent to the parental JEV strain. Simultaneously performed ELISAs using the whole virus antigen and newly developed antigen gave comparable results for JE positive and negative samples, which established the utility of developed JEV E-VLP as an antigen. Reduced cross-reactivity and increased specificity were observed when tested with dual positive sera for anti-JEV and DENV antibodies. These findings confirm the efficiency and reliability of newly developed recombinant E-VLP antigen expressed by the BHK-IE6 cell clone as an antigen in serodiagnostic assays. The implementation and progress in developing cross-reactivity-reduced antigens would improve serodiagnosis and disease burden estimates of flavivirus infection.
Key points
• pcDNA3.1/JE-Sig-prM-E plasmid transfected BHK-21 cells stably express VLPs.
• Sodium butyrate induction enhanced the extracellular expression of VLPs.
• Application of JEV-E VLPs increases the specificity of JE IgM ELISA.
Publisher
Springer Berlin Heidelberg,Springer,Springer Nature B.V
Subject
/ Analysis
/ Animals
/ Antigens
/ Assaying
/ Biomedical and Life Sciences
/ Biotechnologically Relevant Enzymes and Proteins
/ Cloning
/ Encephalitis Virus, Japanese - genetics
/ Encephalitis, Japanese - diagnosis
/ Enzyme-linked immunosorbent assay
/ Enzyme-Linked Immunosorbent Assay - methods
/ Female
/ genes
/ Genotype
/ India
/ Mammals
/ Methods
/ Microbial Genetics and Genomics
/ Plasmids
/ Viremia
/ Viruses
