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DENGUE DIAGNOSTICS
DENGUE DIAGNOSTICS
The Right Test at the Right Time
for the Right Group

edited by
Shamala Devi Sekaran
Published by
Jenny Stanford Publishing Pte. Ltd.
101 Thomson Road
#06-01, United Square
Singapore 307591

Email: [email protected]
Web: www.jennystanford.com

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library.

Dengue Diagnostics: The Right Test at the Right Time for the
Right Group
Copyright © 2024 by Jenny Stanford Publishing Pte. Ltd.
All rights reserved. This book, or parts thereof, may not be reproduced in any form
or by any means, electronic or mechanical, including photocopying, recording
or any information storage and retrieval system now known or to be invented,
without written permission from the publisher.

For photocopying of material in this volume, please pay a copying fee through
the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923,
USA. In this case permission to photocopy is not required from the publisher.

ISBN 978-981-4968-97-3 (Hardcover)

ISBN 978-1-032-66956-4 (eBook)
 Contents

Preface vii

1. Introduction to Dengue and Issues with

Current Diagnostics 1
Shamala Devi Sekaran, Rishya Manikam,
and Gaythri Thergarajan
1.1 Current Epidemiology 2
1.2 Diagnosis 5
1.3 Immune Responses to Dengue 6
1.4 Markers and Problems 8

2. Traditional and Current Diagnosis 13

Wang Seok Mui and Chandramathi Samudi Raju
2.1 Virus Isolation 15
2.1.1 Intracerebral Injection into Mice 15
2.1.2 Inoculation of Specimen into
Mosquitoes 16
2.1.3 Viral Culture Using Mosquito or
Mammalian Cell Line 17
2.2 Detection of Dengue-Specific Antigens and
Antibodies 18
2.2.1 Antigen Detection 18
2.2.1.1 Non-structural protein
(NS1 & NS5)-based assay 18
2.2.2 Antibody Detection 20
2.2.2.1 Hemagglutination
inhibition test 20
2.2.2.2 Envelope/membrane
(E/M)-antigen ELISA
(both IgM and IgG) 22
2.2.2.3 Rapid diagnostic tests 23
2.3 Viral Nucleic Acid Detection 23
2.3.1 Polymerase Chain Reaction (PCR) 23
2.3.2 Nucleic Acid Sequence‒Based
Amplification (NASBA) 24
vi Contents

2.3.3 Loop-Mediated Isothermal

Amplification (LAMP) 25
2.4 Diagnostic Limitation 26
2.5 Future Directions 27

3. Newer Diagnostics for Dengue Disease 35

Crystale Lim Siew Ying, Nur Haliza Hassan,
and Shamala Devi Sekaran
3.1 Biosensors in Dengue Diagnosis 35
3.2 Biosensor Platform 42
3.2.1 NS1 Antigen/Antibody Detection 42
3.2.2 Detection of Anti-Dengue IgM
Antibodies 43
3.2.3 Detection of Dengue Viral Genome 44
3.3 Limitations 45

4. Search for Newer Markers for Dengue Disease 51

Yong Yean Kong, Tan Hong Yen, Shamala Devi Sekaran,
and Rishya Manikam
4.1 Introduction 51
4.2 Viral Markers and Their Association with
Dengue Severity 52
4.3 Clinical Markers of Severe Dengue 55
4.4 Markers of Immune Activation and
Severe Dengue Disease 56
4.5 Cytokines in Dengue Infection 64
4.6 Metabolites Released During a Dengue Infection 72
4.7 Markers of Endothelial Activation and
Coagulopathy 74
4.8 Limitations 77
4.9 Research Aspects Needed 78

5. Research and Recommendations 91

Shamala Devi Sekaran
5.1 Research Aspects Needed 92

Selected Protocols 95

Index 103
 Preface

Dengue is a mosquito-borne viral infection that affects millions of

people worldwide every year. It can cause a range of symptoms,
from mild fever and rash to severe hemorrhagic fever and shock.
Dengue is a major public health problem, especially in tropical and
subtropical regions, where it poses a significant burden on health
systems and economies.
The diagnosis of dengue is crucial for the timely management
and treatment of patients, as well as for the prevention and
control of outbreaks. The diagnosis of dengue is challenging due
to the complexity of the disease, the diversity of the virus, and
the limitations of the available diagnostic methods. The clinical
manifestations are often nonspecific and overlap with other febrile
illnesses. Moreover, the laboratory confirmation of dengue requires
the detection of either the virus, its antigens, or its antibodies,
which have different sensitivities and specificities at different stages
of the infection. Therefore, there is a need for accurate, rapid, and
affordable diagnostic tests that can be used in various settings and
scenarios.
This book is based on my extensive experience and research in
the field of dengue diagnostics as well as that of the experts and
collaborators who have contributed to the chapters. It provides a
comprehensive overview of the current state of the art in dengue
diagnostics, covering the principles, techniques, applications, and
challenges of various methods. It also discusses future directions
and perspectives for the development and improvement of dengue
diagnostics.
The book is intended for researchers, clinicians, public health
professionals, students, and anyone interested in learning more
about dengue diagnostics, and I hope it will be a valuable resource
and reference for them and will inspire further research and
innovation in this important area of global health.
Shamala Devi Sekaran
Summer 2023
Chapter 1

Introduction to Dengue and Issues

with Current Diagnostics

Shamala Devi Sekaran,a Rishya Manikam,b

and Gaythri Thergarajana
aFacultyof Applied Sciences, UCSI University Kuala Lumpur Campus,
Malaysia
bEmergency and Trauma Centre, University Malaya Medical Centre,

Kuala Lumpur, Malaysia

[email protected]

Historically dengue is said to have been derived from the Swahili

phrase “Ki-denga pepo”, which basically translates to mean cramp-
like seizures caused by an evil spirit. The Swahili word may possibly
originate from the Spanish word “dengue”, which actually describes
the gait of a person suffering from intense bone pain (Hotta, 1952;
Skae, 1902). Today, however, we know dengue as a viral infection
caused by the dengue virus belonging to the family Flaviviridae.
The first record of this illness is recorded in the Chinese medical
encyclopedia during the Jin Dynasty (265–420) and the illness was
referred to as a water poison associated with flying insects. The

Dengue Diagnostics: The Right Test at the Right Time for the Right Group
Edited by Shamala Devi Sekaran
Copyright © 2024 Jenny Stanford Publishing Pte. Ltd.
ISBN 978-981-4968-97-3 (Hardcover), 978-1-032-66956-4 (eBook)
www.jennystanford.com
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