Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology

Scrub typhus (ST), also known as tsutsugamushi disease and caused by rickettsia Orientia tsutsugamushi, is an underestimated fatal epidemic in the Asia-Pacific region, resulting in a million human infections each year. ST is easily misdiagnosed as clinical diagnosis is based on non-specific skin esc...

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Autores principales:	Chih-Chi Liao, Chih-Hsuan Tsai, Huei-Ru Lo, Pey-Ru Lin, Chang-Chi Lin, Yu-Chan Chao
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	baculovirus surface display cell-based ELISA Orientia tsutsugamushi scrub typhus serological diagnosis Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/92f60243ef04481591b8c4ea2a949fa7
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spelling	oai:doaj.org-article:92f60243ef04481591b8c4ea2a949fa72021-11-16T09:50:55ZDevelopment of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology1664-322410.3389/fimmu.2021.761136https://doaj.org/article/92f60243ef04481591b8c4ea2a949fa72021-10-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fimmu.2021.761136/fullhttps://doaj.org/toc/1664-3224Scrub typhus (ST), also known as tsutsugamushi disease and caused by rickettsia Orientia tsutsugamushi, is an underestimated fatal epidemic in the Asia-Pacific region, resulting in a million human infections each year. ST is easily misdiagnosed as clinical diagnosis is based on non-specific skin eschar and flu-like symptoms. Thus, the lack of accurate, convenient, and low-cost detection methods for ST poses a global health threat. To address this problem, we adopted baculovirus surface-display technology to express three variants of TSA56, the major membrane antigen of O. tsutsugamushi, as well as the passenger domain of ScaC (ScaC-PD), on insect Sf21 cell surfaces rather than biosafety level 3 bacteria in an enzyme-linked immunosorbent assay (ELISA). Recombinant TSA56 and ScaC-PD were all properly expressed and displayed on Sf21 cells. Our cell-based ELISA comprising the four antigen-displaying cell types interacted with monoclonal antibodies as well as serum samples from ST-positive field-caught rats. This cell-based ELISA presented high accuracy (96.3%), sensitivity (98.6%), and specificity (84.6%) when tested against the ST-positive rat sera. Results of a pilot study using human sera were also highly consistent with the results of immunofluorescence analyses. By adopting this approach, we circumvented complex purification and refolding processes required to generate recombinant O. tsutsugamushi antigens and reduced the need for expensive equipment and extensively trained operators. Thus, our system has the potential to become a widely used serological platform for diagnosing ST.Chih-Chi LiaoChih-Hsuan TsaiHuei-Ru LoPey-Ru LinChang-Chi LinYu-Chan ChaoYu-Chan ChaoYu-Chan ChaoYu-Chan ChaoFrontiers Media S.A.articlebaculovirus surface displaycell-based ELISAOrientia tsutsugamushiscrub typhusserological diagnosisImmunologic diseases. AllergyRC581-607ENFrontiers in Immunology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	baculovirus surface display cell-based ELISA Orientia tsutsugamushi scrub typhus serological diagnosis Immunologic diseases. Allergy RC581-607
spellingShingle	baculovirus surface display cell-based ELISA Orientia tsutsugamushi scrub typhus serological diagnosis Immunologic diseases. Allergy RC581-607 Chih-Chi Liao Chih-Hsuan Tsai Huei-Ru Lo Pey-Ru Lin Chang-Chi Lin Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
description	Scrub typhus (ST), also known as tsutsugamushi disease and caused by rickettsia Orientia tsutsugamushi, is an underestimated fatal epidemic in the Asia-Pacific region, resulting in a million human infections each year. ST is easily misdiagnosed as clinical diagnosis is based on non-specific skin eschar and flu-like symptoms. Thus, the lack of accurate, convenient, and low-cost detection methods for ST poses a global health threat. To address this problem, we adopted baculovirus surface-display technology to express three variants of TSA56, the major membrane antigen of O. tsutsugamushi, as well as the passenger domain of ScaC (ScaC-PD), on insect Sf21 cell surfaces rather than biosafety level 3 bacteria in an enzyme-linked immunosorbent assay (ELISA). Recombinant TSA56 and ScaC-PD were all properly expressed and displayed on Sf21 cells. Our cell-based ELISA comprising the four antigen-displaying cell types interacted with monoclonal antibodies as well as serum samples from ST-positive field-caught rats. This cell-based ELISA presented high accuracy (96.3%), sensitivity (98.6%), and specificity (84.6%) when tested against the ST-positive rat sera. Results of a pilot study using human sera were also highly consistent with the results of immunofluorescence analyses. By adopting this approach, we circumvented complex purification and refolding processes required to generate recombinant O. tsutsugamushi antigens and reduced the need for expensive equipment and extensively trained operators. Thus, our system has the potential to become a widely used serological platform for diagnosing ST.
format	article
author	Chih-Chi Liao Chih-Hsuan Tsai Huei-Ru Lo Pey-Ru Lin Chang-Chi Lin Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao
author_facet	Chih-Chi Liao Chih-Hsuan Tsai Huei-Ru Lo Pey-Ru Lin Chang-Chi Lin Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao Yu-Chan Chao
author_sort	Chih-Chi Liao
title	Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
title_short	Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
title_full	Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
title_fullStr	Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
title_full_unstemmed	Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology
title_sort	development of a scrub typhus diagnostic platform incorporating cell-surface display technology
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/92f60243ef04481591b8c4ea2a949fa7
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Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology

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