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

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S127
reconstituted human airway epithelia (MucilAirTM). Differentiated
tissues were infected in parallel with clinically relevant strains of
rhinovirus (A16, A49, A55, B48, C8, C15), respiratory enterovirus
(EV68), influenza virus (H3N2) and corona virus (OC43). For each
virus, replication kinetics, cell tropism, impact of the virus on tissue
integrity and cilia function were assessed.
Development and use of anti-viral drugs are one of the priorities
for major pharmaceutical companies. As proof-of-concept for drug
screening, the efficacy of Rupintruvir and Oseltamivir were tested
in MucilAirTM. Rupintruvir efficiently inhibited the replication of
HRV-A16 and HRV-C15 in a dose and time dependent manner (up
to 99% inhibition). Interestingly, (i) Oseltamivir reduced the replica-
tion of H1N1 and H3N2 and restored the impaired barrier function
monitored by Trans-Epithelial Electrical Resistance and (ii) Rupin-
tivir restored the mucociliary clearance impaired by EV68 (7 m/s
for the Mock up to 40 m/s for the Rupintrivir treatment at 50 nM
at 96 h post innoculation).
These results demonstrated that MucilAirTM is a robust, reliable
and relevant tool for antiviral drug development.
Abstract no: 346
Presentation at ESCV 2016: Poster 214
Study on immunological characteristics of
monoclonal antibodies produced against the
Kazakhstan isolates of influenza A(H1N1) virus
N.G. Klivleyeva∗, T.I. Glebova, M.G. Shamenova
Institute of Microbiology and Virology, Kazakhstan
The main economic and social damage resulting from infec-
tious diseases throughout the world is caused by acute respiratory
viral infections and influenza. In recent years, the epidemic pro-
cess is characterized by co-circulation of influenza virus subtypes
A(H1N1), A(H3N2) and type B. In determining the etiology of
viral infection serological analysis is one of the fundamental com-
ponents. Monoclonal antibodies (MAbs) permit to dramatically
increase the specificity and sensitivity of diagnostic techniques
for the detection of viral antigens. Immunological characteristics
of MAbs produced against the Kazakhstan isolates of influenza
A(H1N1) virus were studied with immunofluorescence, HAI and
microneutralization assays. Immunofluorescence testing revealed
that MAbs are specific against homologous and related antigens,
and identified them in the form of distinct granular fluorescence
before the conjugate dilutions of 1:80–1:160. It was found that
MAbs in HAI assay revealed a wide range of responses and in high
titres (1:160–1:10240) inhibited the hemagglutinating activity of
the homologous and related reference and Kazakhstan influenza
viruses and did not react with the heterologous A(H3N2) and type
B viruses. In microneutralization assay MAbs neutralized influenza
A(H1N1) viruses and did not react with influenza viruses A(H3N2)
and type B. Thereby, the similar spectra of MAb reactivity against
A/H1N1 viruses indicate the presence of antigenic determinants
in the HA composition of all the investigated viruses, that allows
to recommend the resulting MAbs for differentiation of A(H1N1)
viruses from the seasonal A(H3N2) and type B strains.
Abstract no: 38
Presentation at ESCV 2016: Poster 215
Genetic diversity and characteristics of porcine
reproductive and respiratory syndrome virus in
the area of Korea from 2013 to 2015
I.O. Ouh, J.E. Yu, H. Kang, J. Lee, S.E. Choe, I.S. Cho,
S.H. Cha ∗
Viral Disease Division, Animal and Plant Quarantine
Agency, Gimcheon 39660, Republic of Korea
Introduction: Porcine reproductive and respiratory syndrome
virus (PRRSV) is rapidly gaining importance as one of the most
economically significant diseases in swine worldwide. PRRSV is an
enveloped positive single-stranded RNA virus that can be divided
into two different genotypes, the European genotype (type 1) and
the North American genotype (type 2). The genome of PRRSV is
approximately 15 kb in length and contains at least 20 open read-
ing frames (ORFs). ORF5, encoding GP5, is one of the most variable
regions of the PRRSV genome, and often used to examine genetic
diversity and monitor evolution of PRRSV. In this study, the recent
isolates in the field were evaluated for genetic variation based on
ORF5 nucleotide and amino acid sequence.
Materials and methods: Lung and serum samples were col-
lected from 541 pig farms in nationwide where clinical symptoms
had been observed in 2013–2015. Total RNA was extracted from
serum and lung using an RNeasy mini Kit (QIAGEN) according to
the manufacturer’s protocol. To obtain sequences of the complete
ORF5, reverse transcriptase-polymerase chain reaction (RT-PCR)
was carried out using the One Step RT-PCR Kit (QIAGEN) and PRRSV
primer sets derived from sequences of the ORF4-6 of the LV and
VR-2332 strain, respectively. Multiple sequence alignments and
phylogenetic trees were carried out using CLC Main Workbench
7.0.3 and Mega 6 program. Bootstrap values were calculated on
1000 replicates of the alignments to assess the confidence limits of
the branching.
Results: To more totally understand the genetic diversity and
characteristics of PRRSV in the area of Korea, we analyzed the open
reading frame (ORF) 5 sequences of 323 (type 1) and 269 (type 2) of
PRRSV from2013 to 2015. The results showed that both types 1 and
2 have been circulating in Korea pig farms and that the regional rate
of infection was more prevalent in Gyeongsangnam-do province
in Korea. Type 1 PRRSVs from Korea are clustered in subtype 1,
subgroup A, B, and C. Type 2 PRRSVs are classified in lineage 1, 4,
5 and new Korea subgroup A, B. Recently, the genetics of type 2
PRRSVs in Korea have become unique regional characteristics in
Gyeongsangnam-do. Recently, the genetics of PRRSVs in Asia have
become more diverse. Although the genetics of type 2 PRRSV in
Korea have unique regional characteristics in Gyeongsangnam-do,
the genetics of PRRSV in Asia have become more diverse.
Conclusions: This study of PRRSV indifferent geographical areas
should be performed regularly to monitor field isolates. This would
provide annual genetic information for PRRS control and vaccine
selection and/or renewal
Reference
[1] E.J. Choi, et al., Genetic diversity of porcine reproductive and respiratory
syndrome virus in Korea, J. Vet. Sci. 14 (2) (2013) 115–124.
[2] S.H. Kim, et al., A molecular analysis of European porcine reproductive and
respiratory syndrome virus isolated in South Korea, Vet. Microbiol. 143 (2010)
394–400.