Journal of Clinical Virology - Volume 82 - Supplement 1 - September 2016

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S105
Method: Laboratory data for patient samples sent for
enterovirus culture from 2008–2015, stored on the laboratory
information system (LIS), were retrieved and analysed. Laboratory
data comprised the test result, age, and gender of the patients.
Results: Out of 1890 samples comprising stool, rectal swabs,
throat swabs, oral ulcer swabs, autopsied heart and intestinal tis-
sues, 173 enterovirus isolates were obtained. Therewere 21 (28.8%)
isolates from patients aged <2 months, 101 (58.4%) isolates from
patients aged 2months to <3 years, 31 (17.9%) isolates frompatients
aged 3 to <7 years, 15 (8.7%) isolates from patients aged 7 to <15
years, and 5 (2.9%) isolates frompatients 15 years and older. Amale
predominance (65.9% male, 34.1% female) was observed. There
were 22 enterovirus serotypes identified. Majority (74%) of the iso-
lates were members of the species human enterovirus A, which
include the serotypes EV-71 (55, 33%), CV-A6 (31, 19%), CV-A10 (17,
10%), and CV-A16 (13, 8%), which are associated with HFMD. The
remaining (26%) were members of the species human enterovirus
B, which include E19 (9, 5%) and CV-B5 (8, 5%).
Out of 1269 cerebrospinal fluid (CSF) and autopsied brain tis-
sue examined, 16 enterovirus isolates were obtained from patients
aged 11 days to 40 years. Therewere 12 (75%) isolates frompatients
aged <3 months. No gender predominance (50% male, 50% female)
was observed. There were 10 enterovirus serotypes identified. The
predominant serotype was CV-B5 with 5 isolates (31.3%).
Conclusion: The predominant serotypes isolated are those com-
monly associated with HFMD, due to the laboratory’s participation
in Singapore’s Ministry of Health HFMD surveillance programme.
Thus, unlike reports from Marseille and the United States where
HEV-B species account for the majority of isolates, this study
shows a predomination of HEV-A species. The largest number
of isolates was obtained from the age group eligible for enrol-
ment in infant/child care centres prior to entering pre-school (2
months to <3 years). This reinforces the importance of infec-
tion control in care centres with infants and young children to
minimise the spread of infectious diseases. Currently, Singapore’s
enterovirus surveillance only includes HFMD, poliovirus and
enteroviral encephalitis. It might be beneficial to conduct routine
surveillance of enteroviruses, similar to the National Enterovirus
Surveillance System (NESS) in the United States, to document dis-
ease associations, and provide samples of new viruses for study.
Abstract no: 147
Presentation at ESCV 2016: Poster 170
Prevalence of enterovirus in patients with
meningitis: 2007–2015
Candan Cicek ∗, Imre Altuglu , Ruchan Sertoz ,
Aysin Zeytinoglu , Selda Erensoy ,
Husnu Pullukcu , Eylem Ulas Saz
1 Ege University, Faculty of Medicine, Department of
Medical Microbiology, Izmir, Turkey
2 Ege University, Faculty of Medicine, Department of
Infectious Diseases, Izmir, Turkey
3 Ege University, Faculty of Medicine, Department of
Pediatrics, Izmir, Turkey
Objectives: Viruses are the major cause of the acute meningitis.
Nonpolio enteroviruses (%80–95) account for most of the aseptic
meningitis cases for which an etiologic agent is identified. The aim
of this study was to establish the prevalence of enteroviruses in
patients with aseptic meningitis in our region.
Methods: Between January 2007 and December 2015, cere-
brospinal fluid (CSF) specimens were collected from 576 [250
(43.4%) female, 376 (56.6%) male] paediatric and adult patients
who admitted with meningitis symptoms in Ege University Fac-
ulty of Medicine Hospital. The age range of patients is between
21 days to 90 years (median: 11 years). Of the 576 patients, 375
(65.1%) were paediatric, and 201 (34.9%) were adult patients. Shell
vial cell culture method was performed for all the specimens sent
to cell culture laboratory. After specimens were vortexed, three
shell vials were prepared for each patient [Human laryngeal car-
cinoma (HEp-2), embryonic rhabdomyosarcoma (RD), and African
green monkey kidney (Vero) cells line]. Each vial was inoculated
with 0.5ml CSF for the recovery of enteroviruses. The vials were
centrifuged at 700 ×g for 30min at 25 ◦C and incubated at 37 ◦C
for 1 h. Supernatants were aspirated from each vial. Subsequently,
1ml isolation medium containing Eagles MEM supplemented with
2% FCS and antibiotics was added to the vials containing HEp-2, RD,
and Vero cells. Then, the vials were incubated in moist chamber at
37 ◦C in a 5% CO2 atmosphere for 48 h. Coverslips were fixed and
stained with a fluorescein isothiocyanate (FITC) labelled polyclonal
antibody specific for enteroviruses (Pan-Enterovirus Blend, Chemi-
con International, USA) according to the manufacturer’s protocol.
The coverslips which had one or more fluorescing inclusions bodies
were considered as positive.
Results: In 576 patients, 35 (6.1%) patients were positive for
enteroviruses. Of the 35 patients (17 female, 18 male), 10 were
adult (median: 35 years) and 25 were paediatric (median: 8 years)
patients. All the patients that are enteroviruses positive had the
clinical diagnosis of aseptic meningitis except one patient who has
encephalopathy.
Conclusion: Most of patients (71.4%) which enterovirus has
been detected were paediatric patients. Aseptic meningitis was
the most common clinical diagnosis. The overall prevalence of
enteroviruses was 6.1% in our region.
Abstract no: 163
Presentation at ESCV 2016: Poster 171
Controlling the quality of diagnostic PCR assays
Kathryn Doris ∗, Sarah Kempster,
Cristina Santirso-Margaretto, Graham Prescott,
Clare Morris, Neil Almond, Rob Anderson
NIBSC, United Kingdom
The aim of infectious disease diagnostics is the early detection
of infection to allow for the administration of appropriate ther-
apy. Quantitative real time PCR (qPCR) has revolutionised the fields
of pathogen detection by offering faster, more accurate and more
sensitive results than traditional methods. However qPCR is not
without problems, principally of assay performance.
It is essential that laboratories monitor the day-to-day per-
formance of an assay, specifically with respect to reproducibility
and repeatability, so that the laboratory may be confident that the
results generated by the assay lead to appropriate diagnoses. Fre-
quently laboratories do not run external standards which would
allow for easy performance monitoring. This is often due to the
unavailability of appropriate controls and an unavailability of a
means of monitoring assay results. NIBSC have sought to address
these issues in two ways.
First, NIBSC have produced a series of CE-marked multiplex
run control reagents for qPCR. These reagents are formulated with
intact viruses and therefore can be used as a whole process con-
trols. The concentrations of all of the analytes in the reagents are
set at a level designed to be challenging to an assay but to be con-
sistently detected as lowpositives. The controls are available freeze