Disease: Influenza (Flu) Gene of interest: “H275Y”.

1. Dr. Obumneke Amadi-Onuoha Scripts
Disease: Influenza (Flu)
Gene of interest: “H275Y”.
ResearchQuestion:
Would the presence of the changes in H275Y neuraminidase gene be a reliable predictor of
reduced susceptibility and antiviral resistance of influenza infection (Flu Virus) in affected
individuals or not?
Background:
There are four types of influenza viruses (A to D). Human influenza A and B viruses cause
seasonal epidemics in the United States. Spontaneously, the emergence of a new and very
different influenza A virus occurs that infect people and can cause an influenza pandemic (CDC,
2017). Influenza A viruses are subdivided into two based on two proteins on the surface of the
virus: the hemagglutinin (H) and the neuraminidase (N) and are broken down into different
strains. The existing subtypes of influenza A viruses found in people are influenza A (H1N1) and
influenza A (H3N2) viruses. In the year 2009, a new influenza A (H1N1) virus emerged to
cause illness in people and a caused the first influenza pandemic (CDC, 2017), this virus was
very different from the human influenza A (H1N1) viruses spreading at that time. The influenza
A(H1N1) pdm09 has now replaced the H1N1 virus that was previously spreading in humans, the
risk of a neuraminidase inhibitor-resistant strain developing is of concern (CDC, 2017). The
resistance in influenza A(H1N1) pdm09 has been described to be associated with the histidine-
to-tyrosine (H275Y) mutation in the neuraminidase (NA) gene (Pawestri, Nugraha, Hariastuti, &
Setiawaty, 2018, p.1-2). This virus can become a major global, clinical and public health issue.
It is important for the constant monitoring for NA inhibitor resistant viruses to predict any

2. increases in the number of resistant viruses since NA inhibitors are used largely for treatment
and outbreak interventions (Pawestri, Nugraha, Hariastuti, & Setiawaty, 2018, p.1-2).
Short justification: Influenza (flu) is a contagious respiratory illness caused by influenza
viruses. Influenza virus A (HINI) occasionally change their strain in the course of flue seasons,
the reason being that flu virus replicates. These changes genetic makeup may change the ways
of treatment/therapy adherence and the effectiveness of the antiviral drugs against these viruses
(cdc, 2019). Among the vaccines used in the treatment for the infection, oseltamivir is the most
commonly prescribed and recommended in the United States. A gene the “H275Y” is conferred
to resist oseltamivir in 2009 H1N1 flu viruses (CDC, 2019). H275Y substitution in the active
site of the NA-1 gene indicates’ reduced binding affinity of the neuraminidase inhibitor
oseltamivir (Hindiyeh, 2010, p.1884). Detection of reduced susceptibility and antiviral resistance
involves local and advanced laboratory tests, such as specific functional assays and molecular
techniques (sequencing and pyrosequencing) to identify the genetic changes linked with reduced
antiviral susceptibility (cdc, 2019). According to Grund et al. (2015), to access spread and
clinical impact of these viruses, it is vital to conduct antiviral susceptibility testing and reporting
of viruses carrying amino acid substitutions in order to confer their antiviral drug resistance
strength (Grund, et al., 2015).
The purpose of this study is to investigate the genomic aspect of the viral infections H1N1pdm09
strain and the mutation in neuroaminidase (NA) gene H275Y to confirm its responsibility for
reduced susceptible and antiviral drug resistance in affected people and examine if there is a
difference in the gene expression of the virus infection in 2 groups: normal and affected people
(children, adults and immunocompromised). The outcome of the study will give a greater
insight of the gene responsibility for reduced susceptibility and antiviral resistance that may

3. guide clinicians as a reliable predictor or biomarkers to identify presence of the virus for support
towards efficient and effective vaccine productions, intervention & treatment. It may help
providers towards diagnosis/planning during spontaneous change in flu season. Also, the study
will confirm previous research knowledge of the gene capability to reliably predict changes in
H275Y mutation in the neuraminidase (NA) gene to monitor a potential antiviral developing
inhibitor-resistant strain.
Hypothesis: Presence of either one of the 3 mutations is sufficient to develop a change in gene
expression that are responsible for immune response against influenza virus and vaccine.
Aims:
1) Investigate the functional outcome of each of the mutations - what changes in H275Y
functions occur when each mutation is present
2) To estimate the probability of developing immune response against influenza virus and
vaccine in the presence of all 3 mutations in H275Y.
Summary of the selectedgene:
Gene symbol: H275Y
Gene description: neuraminidase
Locus tag: FLUAVs6gp1
Gene type: protein coding
RefSeq status: PROVISIONAL
Organism: Influenza A virus (A/Puerto Rico/8/1934(H1N1)) (strain: A/Puerto
Rico/8/1934(H1N1))
Lineage: Viruses; ssRNA viruses; ssRNA negative-strand viruses; Orthomyxoviridae;
Influenzavirus A

4. Location: segment: 6
Sequence: NC_002018.1 (21..1385)
Method: Investigating activities of potential promoter DNA, gene polymorphism, and gene
expression in people with and without influenza A (H1N1) infection.
Protocol development:
1. Literature review: Examination of primary research and systematic reviews of H275Y and
oseltamivir-resistant data and link with influenza A(H1N1)pdm09 viruses.
2. Bioinformatics: Collection of genomic data on H275Y and influenza A(H1N1)pdm09 viruses
disease.
Reference
CDC (2019). Influenza (FLU): Antiviral Drug Resistance. Retrieved from
https://www.cdc.gov/flu/treatment/antiviralresistance.htm
CDC (2017). Types of Influenza Viruses. Retrieved from
https://www.cdc.gov/flu/about/viruses/types.htm
Grund, S., Gkioule, C., Termos, T., Pfeifer, N., Kobbe, G., Verheyen, J., & Adams, O. (2015).
Case report Primarily oseltamivir-resistant influenza A (H1N1pdm09) virus evolving into
a multidrug-resistant virus carrying H275Y and I223R neuraminidase substitutions.
Antiviral therapy, 20, 97-100.
Hindiyeh, M., Ram, D., Mandelboim, M., Meningher, T., Hirsh, S., Robinov, J., ... &

5. Mendelson, E. (2010). Rapid detection of influenza A pandemic (H1N1) 2009 virus
neuraminidase resistance mutation H275Y by real-time reverse transcriptase PCR.
Journal of clinical microbiology, 48(5), 1884-1887.
Pawestri, H. A., Nugraha, A. A., Hariastuti, N. I., & Setiawaty, V. (2018). Detection of
neuraminidase inhibitor-resistant influenza A (H1N1) pdm09 viruses obtained from
influenza surveillance in Indonesia. SAGE open medicine, 6, 2050312118818293