Antiviral drug resistance of herpes simplex virus Růžena Stránská

137 Pages·2004·1.39 MB·Dutch

Antiviral drug resistance
of herpes simplex virus

Růžena Stránská
Printed by Febodruk B.V., Enschede

ISBN: 90-393-3609-1
© R. Stránská, Utrecht 2004
All rights reserved
Antiviral drug resistance of herpes simplex virus

Antivirale resistentie van het herpes simplex virus

Rezistence viru herpes simplex vůči virostatikům

(met een samenvatting in het Nederlands)

(se shrnutim v českém jazyce)

Proefschrift
ter verkrijging van de graad van doctor
aan de Universiteit Utrecht
op gezag van de Rector Magnificus, Prof. Dr. W. H. Gispen
ingevolge het besluit van het College voor Promoties
in het openbaar te verdedigen
op vrijdag 20 februari 2004 des ochtends te 10.30 uur

door

Růžena Stránská

geboren op 24 september 1975 te Brandýs nad Labem, Tsjechië
promotor: Prof. Dr. J. Verhoef
co-promotoren: Dr. A. M. van Loon
Dr. R. Schuurman

The experimental work described in this thesis was performed at the Department of Virology,
Eijkman-Winkler Center, University Medical Center Utrecht, The Netherlands and was supported
by a grant ZON 97-1-297 from ZorgOnderzoek Nederland.
The printing of this thesis was financially supported by J. E. Jurriaanse Stichting, Dr. Ir. van de
Laar Stichting, Eijkman Graduate School for Immunology and Infectious Diseases,
GlaxoSmithKline B.V., Oxoid B.V., Roche Diagnostics B.V.
No pessimist ever discovered
the secrets of the stars,
or sailed to uncharted land,
or opened a new doorway for the human spirit.

Helen Keller

Věnováno mým rodičům
Voor mijn ouders

Contents

Chapter 1: Introduction 9

Chapter 2: Routine use of a highly automated and internally controlled
real-time PCR assay for the diagnosis of herpes simplex and
varicella-zoster virus infections
Journal of Clinical Virology. In press 29

Chapter 3: Application of real-time PCR for antiviral drug susceptibility
determination of herpes simplex virus
Antimicrobial Agents and Chemotherapy. 2002, 46: 2943-2947 39

®
Chapter 4: ELVIRA HSV-a yield reduction assay for rapid antiviral
susceptibility testing of herpes simplex virus
Submitted 51

Chapter 5: Survey of acyclovir-resistant herpes simplex virus
in The Netherlands: prevalence and characterization
Submitted 65

Chapter 6: Genotypic and phenotypic characterization of acyclovir-resistant
herpes simplex viruses isolated from hematopoietic stem cell
transplant recipients
Submitted 85

Chapter 7: Sequential switching of DNA polymerase and thymidine kinase
mediated HSV-1 drug resistance in an immunocompromised child
Antiviral Therapy. 2004, 9: 97-104 105

Chapter 8: General discussion 119

Nederlandse samenvatting (Summary in Dutch) 129

Shrnutí v českém jazyce (Summary in Czech) 131

Acknowledgements 133

Curriculum vitae 136

List of publications 137

Chapter 1

Introduction
Chapter 1
Herpesviruses
The family of Herpesviridae comprises a diverse group of large enveloped DNA viruses
of vertebrates. The fundamental characteristic of this virus group is the ability to establish
latent infection and periodically reactivate. Human herpesviruses (HHV) include herpes
simplex virus (HSV) type 1 and 2, varicella-zoster virus (VZV), cytomegalovirus (CMV),
Epstein-Barr virus (EBV), and HHV-6, HHV-7 and HHV-8. Based on the host range,
duration of replication cycle, cytopathology and type of latent infection, herpesviruses are
divided into 3 subfamilies: Alpha-, Beta- and Gammaherpesvirinae.

Alphaherpesviruses
Members of the α-herpesvirinae subfamily are characterized by a short replication cycle,
wide host range and a rapid initial lytic infection of mucocutaneous cells followed by
establishment of neuronal latency. This subfamily comprises 3 human herpesviruses: HSV-1,
HSV-2 and VZV. All three are associated with primary and recurrent infections of
mucocutaneous tissues, which may manifest as severe, persistent and disseminated disease
among immunocompromised patients108. The seroprevalence varies between 50-95% for
HSV-1 and 6-50% for HSV-2 depending on the age, sex, race, marital and socioeconomic
status and geographical location130. About 95% of population above the age of 10 years is
seropositive for VZV108,157.

Pathogenesis of HSV infections
After primary lytic infection in peripheral mucocutaneous tissues, HSV establishes a
lifelong latent ganglionic infection of sensory neurons that innervate the area of the primary
infection. From its state of latency, when no virus replication takes place, the virus
periodically reactivates and travels by axonal transport to the epithelial surface of the
inervated region where it causes a recurrent infection154. Primary as well as recurrent
infections may be associated with clinical symptoms or be asymptomatic, but in both cases
virus shedding occurs and virus transmission is possible.
Primary HSV-1 infections of immunocompetent subjects are usually self limiting, and
clinically manifest as gingivostomatitis, keratoconjuctivitis, cutaneous or genital herpes.
Recurrences lead to the same symptoms but are of lesser severity and of shorter duration,
with exception of HSV encephalitis. HSV-2 causes primary genital herpes, but is also an
important cause of meningoencephalitis and neonatal herpes. In immunocompromised
patients, the above mentioned HSV infections are often more severe and have a protracted
course. In addition, HSV esophagitis, pneumonia, hepatitis or disseminated infection may
occur in these patients.
VZV is the causative agent of chicken pox upon primary infection and can result in herpes
zoster (shingles) in adults or in the disseminated infections in immunocompromised patients.

Laboratory diagnosis of HSV infections
Detection of HSV belongs to the frequent diagnostic procedures in the clinical virology
laboratory. For years, virus isolation by cell culture or rapid shell vial culture combined with
immunoflouorescence detection of virus antigen using type-specific monoclonal antibodies
10