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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S63
83 pathogens were detected among the children attending
the emergency care unit. The most commonly detected was
Rotaviruses (n = 70, 84.3%), followed by adenovirus (n = 8, 9.6%) and
astrovirus (n = 3, 3.6%). One sample was positive for norovirus G2
and one sample was positive for sapovirus. Four coinfections were
detected involving rotavirus and adenovirus (n = 2) and rotavirus
and astrovirus (n = 2).
Nine pathogens were detected among the neonates. The most
commonly detected was rotavirus (n = 4, 44.4%), followed by
sapovirus (n = 4, 44.4%). One sample was positive for adenovirus.
One sample showed co-infection (rotavirus and sapovirus).
The FTD viral gastroenteritis® detected 13 more rotavirus
(p < 0.005) than did the BIOSYNEX Adenovirus-Rotavirus
Combo® . Nine samples were adenovirus positive with the
FTD viral gastroenteritis® and were negative with the BIOSYNEX
Adenovirus–Rotavirus, while three samples gave the opposite
results. Tested with the adenovirus in-house real-time PCR, 6/9
samples were positive (p < 0.05), all the remaining three were
negative.
Conclusion: The FTD viral gastroenteritis® is a very sensitive
and convenient method for detecting multiple gastrointestinal
virological pathogens.
Abstract no: 208
Presentation at ESCV 2016: Poster 84
Comparison of rotavirus frequency and
genotype distribution in Rwanda before and
after vaccine introduction
Maria Andersson ∗, Jean-Claude Kabayiza ,
Magnus Lindh
1 Department of Infectious Diseases, University of
Gothenburg, Gothenburg, Sweden
2 Department of Pediatrics, National university of
Rwanda, Kigali, Rwanda
Acute gastroenteritis is a major reason of disease and death
among children in developing countries. A wide range of viruses,
bacteria and protozoa, with rotavirus as one of the most important
pathogens, can cause infectious diarrhoea. In May 2012, rotavirus
vaccination with pentavalent RotaTeq vaccine was introduced in
the national immunization program in Rwanda. The vaccine cov-
erage in children younger than 1 year of age was 98% in 2014 and
the number of hospital admission for diarrhoea specific to rotavirus
after vaccine introduction has recently been reported to decrease
significantly. Still, the incidence and the genotype distribution of
rotavirus infections in the community are not known. In this work
we compared the frequency of rotavirus together with other diar-
rhoeagenic pathogens, and analysed rotavirus genotypes before
and after the introduction of vaccination.
Faeces samples from children below five years of age, collected
before vaccine introduction between 2010 and 2012 (n = 829) and
after in 2014 (n = 658, 89% vaccinated), were analysed by real-time
PCR targeting a wide range of pathogens. Rotavirus positive sam-
ples were genotyped by a real-time PCR assay targeting rotavirus
VP7 (G1, G2, G3, G4, G9 and G12) and VP4 (P[4], P[6] and P[8]) to
identify the most common genotypes.
The proportion of samples that were of rotavirus positive
was 34% before vaccine introduction. In 2014, the rotavirus was
detected in 30% of vaccinated and 27% of unvaccinated children.
Interestingly, norovirus genogroup II, sapovirus and astrovirus
were significantly (p > 0.0001) more common in 2014, after vaccine
introduction. There were marked shifts in rotavirus genotypes over
time: in 2010, G2P[4] was detected in 87%, G12P[6] in 13%; in 2011,
G9P[8] was detected in 48%, G1P[8] in 30% and G12P[6] in 16%; in
2012, G9P[8] was detected in 68% and G12P[6] in 21%. In 2014, after
vaccine introduction, G12P[8] was found 81% and G4P[8] in 19%,
without difference between vaccinated and unvaccinated children.
The results show no difference in rotavirus frequency after the
introduction of rotavirus vaccine into the immunisation program,
and that rotavirus infection frequencies were similar in vaccinated
and unvaccinated children. Marked changes of the rotavirus geno-
type distribution were observed from year to year also before
the introduction of vaccination in 2012, and therefore genotype
changes after that time point may not be due to vaccination itself.
Abstract no: 230
Presentation at ESCV 2016: Poster 85
Evaluation of VIASURE real-time PCR assays for
detection of rotavirus and norovirus GI and GII
in fecal samples
C. Santiso-Bellón ∗, S. Vila-Vicent , R. Falcón ,
T. Pascual-Martín , J. Buesa
1 University of Valencia, Spain
2 Hospital Clinico Universitario de Valencia, Spain
Introduction: Rotaviruses (RVs) and noroviruses (NoVs) are
the main etiological agents of nonbacterial acute gastroenteritis
(AGE) in both children and adults. A rapid and sensitive detec-
tion is crucial to implement measures to reduce the spread of
gastroenteritis infections. We evaluated the efficacy of the VIA-
SURERotavirus Real Time PCRDetectionKit, theVIASURENorovirus
GI Real Time PCR Detection Kit and the VIASURE NoV GII Real
Time PCR Detection Kit (Certest Biotec) for their diagnostic appli-
cation in a clinical laboratory. The sensitivity and specificity of
the VIASURE Rotavirus assay was compared with those of the
Rotavirus–Adenovirus immunochromatographic (ICG) test (Certest
Biotec), an in-house conventional RT-PCR for rotavirus detection
and the RIDA ®GENE Viral Stool Panel II real-time RT-PCR (R-
Biopharm AG). The performance of the VIASURE Norovirus GI and
GII assays was also compared with an in-house conventional RT-
PCR for norovirus detection and the RIDA® GENE Norovirus I & II
real-time RT-PCR (R-Biopharm AG).
Materials andmethods:Fecal samples fromchildren and adults
with AGE were collected and diluted in PBS to prepare 10% fecal
extracts. The presence of RV was tested in 210 samples and NoV
GI and GII were analyzed in 181 samples. RNA was extracted
from 200 l of fecal suspensions by using the VIASURE RNA/DNA
Extraction Kit (Certest Biotec). The immunochromatographic (ICG)
test for rapid RV detection was performed following the manu-
facturer’s instructions. Conventional RV RT-PCR reactions for VP7,
VP4 and/or VP6 viral genes were carried out according to the pro-
tocols described by the European Rotavirus Network
. A conventional NoV RT-PCR was performed using
primers JV12/JV13 targeting the polymerase gene and eventual
capsid gene amplification. The real-time RT-PCR assays evaluated in
this studywere performed according to themanufacturer’s instruc-
tions using the StepOneTM real-time PCR equipment (Applied
Biosystems) for the VIASURE kits and the ABI 7500 real-time PCR
System (Applied Biosystems) for the RIDAGENE kits. Samples with
a Ct value ≤35 were considered positive.
Results: Regarding RV detection, both ICG and conventional
RT-PCR detected the presence of RV in 34.3% (72/210) samples,
whereas the VIASURE Rotavirus Real Time PCR assay detected 42.4%
(89/210) positive specimens. The RIDAGENE Viral Stool Panel II