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Development of a sensitive immunochromatographic kit using fluorescent silica nanoparticles for rapid serodiagnosis of amebiasis

Published online by Cambridge University Press:  09 May 2018

Hiroshi Tachibana*
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Azumi Kakino
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Makoto Kazama
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan
Meng Feng
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China
Satomi Asai
Affiliation:
Department of Laboratory Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Kazuo Umezawa
Affiliation:
Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Tsukasa Nozaki
Affiliation:
Central Clinical Laboratory, Tokai University Hospital, Isehara, Kanagawa 259-1193, Japan
Takashi Makiuchi
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Takayuki Kamada
Affiliation:
Furukawa Electric Advanced Engineering Co. Ltd., Ichihara, Chiba 290-8555, Japan
Hideki Watanabe
Affiliation:
Furukawa Electric Advanced Engineering Co. Ltd., Ichihara, Chiba 290-8555, Japan
Noriyuki Horiki
Affiliation:
Department of Endoscopy, Mie University School of Medicine, Tsu, Mie 514-8507, Japan Department of Internal Medicine, St. Luke's International Hospital, Tokyo 104-8560, Japan
Xunjia Cheng
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China
Gohta Masuda
Affiliation:
Department of Infectious Diseases, Tokyo Metropolitan Cancer and Infectious Diseases Center, Komagome Hospital, Tokyo 113-8677, Japan
*
Author for correspondence: Hiroshi Tachibana, E-mail: [email protected]

Abstract

We have previously shown that the C-terminal region of the intermediate subunit of Entamoeba histolytica galactose- and N-acetyl-D-galactosamine-inhibitable lectin (C-Igl) is a useful antigen for serodiagnosis of amebiasis. An immunochromatographic kit was developed using fluorescent silica nanoparticles coated with C-Igl prepared in Escherichia coli. Samples for examination were added to the freeze-dried particles and then applied to the immunochromatographic device, in which a test line on the membrane was also coated with C-Igl. Fluorescent intensity was measured using a hand-held reader. In an evaluation of the kit using a human monoclonal antibody, the minimum amount of C-Igl specific antibody showing positive results was 100 pg. In the evaluation of serum samples with different antibody titers in indirect immunofluorescent antibody tests in the kit, 20 µL of serum was sufficient to obtain positive results at 30 min. Serum samples from symptomatic patients with amebic colitis and amebic liver abscess and those from asymptomatic E. histolytica-cyst carriers showed positive results in the kit. Based on evaluation using sera from healthy controls and patients with other infectious diseases, the sensitivity and specificity of the kit were 100 and 97.6%, respectively. Therefore, we conclude that the newly developed kit is useful for rapid serodiagnosis of amebiasis.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

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