Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes

Dengue is an important public health problem worldwide, with India contributing nearly a third of global dengue disease burden. The measurement of neutralizing antibody responses is critical for understanding dengue pathophysiology, vaccine development and evaluation. Historically, dengue virus neut...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Pragati Sharma, Kaustuv Nayak, Elluri Seetharami Reddy, Humaira Farooqi, Kaja Murali-Krishna, Anmol Chandele
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	dengue flowcytometry neutralization focus reduction neutralizing tests Medicine R
Acceso en línea:	https://doaj.org/article/3a3a9e99dd3a4c8ba688e71b17392a0b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:3a3a9e99dd3a4c8ba688e71b17392a0b
record_format	dspace
spelling	oai:doaj.org-article:3a3a9e99dd3a4c8ba688e71b17392a0b2021-11-25T19:11:27ZOptimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes10.3390/vaccines91113392076-393Xhttps://doaj.org/article/3a3a9e99dd3a4c8ba688e71b17392a0b2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-393X/9/11/1339https://doaj.org/toc/2076-393XDengue is an important public health problem worldwide, with India contributing nearly a third of global dengue disease burden. The measurement of neutralizing antibody responses is critical for understanding dengue pathophysiology, vaccine development and evaluation. Historically, dengue virus neutralization titers were measured using plaque reduction neutralization tests (PRNTs), which were later adapted to focus reduction neutralization tests (FRNTs). Given the slow and laborious nature of both these assays, there has been interest in adapting a high-throughput flow cytometry based neutralization assay. However, flow cytometry based assays typically underestimate neutralization titers, and in situations where the titers are low they can even fail to detect neutralization activity. In this study, by evaluating graded numbers of input Vero cell numbers and viral inoculum, we optimized the flow cytometry based neutralization assay in such a way that it is sensitive and scores titers that are in concordance with focus reduction neutralization tests for each of the four dengue virus serotypes (<i>p</i> < 0.0001). Given that dengue is a global public health concern, and several research groups are making efforts to understand its pathophysiology and accelerate vaccine development and evaluation both in India and worldwide, our findings have timely significance for facilitating these efforts.Pragati SharmaKaustuv NayakElluri Seetharami ReddyHumaira FarooqiKaja Murali-KrishnaAnmol ChandeleMDPI AGarticledengueflowcytometryneutralizationfocus reduction neutralizing testsMedicineRENVaccines, Vol 9, Iss 1339, p 1339 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	dengue flowcytometry neutralization focus reduction neutralizing tests Medicine R
spellingShingle	dengue flowcytometry neutralization focus reduction neutralizing tests Medicine R Pragati Sharma Kaustuv Nayak Elluri Seetharami Reddy Humaira Farooqi Kaja Murali-Krishna Anmol Chandele Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
description	Dengue is an important public health problem worldwide, with India contributing nearly a third of global dengue disease burden. The measurement of neutralizing antibody responses is critical for understanding dengue pathophysiology, vaccine development and evaluation. Historically, dengue virus neutralization titers were measured using plaque reduction neutralization tests (PRNTs), which were later adapted to focus reduction neutralization tests (FRNTs). Given the slow and laborious nature of both these assays, there has been interest in adapting a high-throughput flow cytometry based neutralization assay. However, flow cytometry based assays typically underestimate neutralization titers, and in situations where the titers are low they can even fail to detect neutralization activity. In this study, by evaluating graded numbers of input Vero cell numbers and viral inoculum, we optimized the flow cytometry based neutralization assay in such a way that it is sensitive and scores titers that are in concordance with focus reduction neutralization tests for each of the four dengue virus serotypes (<i>p</i> < 0.0001). Given that dengue is a global public health concern, and several research groups are making efforts to understand its pathophysiology and accelerate vaccine development and evaluation both in India and worldwide, our findings have timely significance for facilitating these efforts.
format	article
author	Pragati Sharma Kaustuv Nayak Elluri Seetharami Reddy Humaira Farooqi Kaja Murali-Krishna Anmol Chandele
author_facet	Pragati Sharma Kaustuv Nayak Elluri Seetharami Reddy Humaira Farooqi Kaja Murali-Krishna Anmol Chandele
author_sort	Pragati Sharma
title	Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
title_short	Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
title_full	Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
title_fullStr	Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
title_full_unstemmed	Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes
title_sort	optimization of flow-cytometry based assay for measuring neutralizing antibody responses against each of the four dengue virus serotypes
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/3a3a9e99dd3a4c8ba688e71b17392a0b
work_keys_str_mv	AT pragatisharma optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes AT kaustuvnayak optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes AT elluriseetharamireddy optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes AT humairafarooqi optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes AT kajamuralikrishna optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes AT anmolchandele optimizationofflowcytometrybasedassayformeasuringneutralizingantibodyresponsesagainsteachofthefourdenguevirusserotypes
_version_	1718410148033069056

Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes

Ejemplares similares