Amantadine resistance of clade 2.3.2 H5N1 Avian Influenza Virus from Waterfowl in Indonesia

Dyah Ayu Hewajuli,I Wayan Teguh Wibawan,Nlp Indi Dharmayanti

doi:10.14334/jitv.v26i3.2764

Dyah Ayu Hewajuli, I Wayan Teguh Wibawan + Show 1 more

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https://doi.org/10.14334/jitv.v26i3.2764

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Abstract

The objective of this research was to know the sensitivity of H5N1 clade 2.3.2 AIV from Indonesia to antiviral drug (amantadine) through molecular and in vitro tests. The study was conducted by virus isolation and identification, nucleotide analysis, and susceptibility to the amantadine hydrocloride in MDCK cells. The study result represented that the mean EID50 isolates of H5N1 clade 2.3.2 AIV was determined of &gt;108 EID50/ml. The analysis of phylogenetic tree of M2 gene from six viruses of H5N1 clade 2.3.2 AIV from Indonesia were closed with H5N1 clade 2.3.2 AIV avian influenza viruses from Vietnam, China, Hongkong. The substitution of M2 protein (V27I) was identified in six isolates H5N1 clade 2.3.2 AIV isolated from Indonesia. Avian influenza of clade 2.3.2 H5N1 subtype from Indonesia produced the formation of CPE and the positive HA reaction with non-toxic concentration of amantadine hydrochloride in MDCK cells. The result of genetic analysis of M2 gene for amantadine resistance was related with the results of HA test and the formation of CPE in MDCK cells. These results established that amantadine resistance have been identified in H5N1 clade 2.3.2 AIV viruses isolated from Indonesia

Highlights

Pathogenic avian influenza (HPAI) virus has been causing significant outbreaks in poultry in Indonesia since 2003 (Wibawa et al 2014; Dharmayanti et al 2014)
The purpose of this research is to study the amantadine susceptibility among avian influenza of 2.3.2 clade isolated from Indonesia based on identification of genetic markers of amantadine resistance and phenotypic analysis in MDCK cells
The presence of H5N1 subtype avian influenza were confirmed by Reverse Transcriptation Polymerase Chain Reaction (RT-PCR) methods with the hemagglutination agglutination (HA) primer sequences of RT PCR method referred to the primers design of Dharmayanti et al 2016 and Lee et al 2001; the NA primer sequence (Wright et al 1995), the Matrix primer sequence (Hoffmann et al 2001)

Summary

Introduction

Pathogenic avian influenza (HPAI) virus has been causing significant outbreaks in poultry in Indonesia since 2003 (Wibawa et al 2014; Dharmayanti et al 2014). The high mortality of ducks caused influenza A (H5N1) viruses of clade 2.3.2.1 were reported in Java Island, Indonesia, at September 2012 (Dharmayanti et al 2014). Transmission of clade 2.1 H5N1 subtype to humans have been discovered in Indonesia while infection of clade 2.3.2 H5N1 subtype in humans have not been officially reported in Indonesia. In Indonesia, the H5N1 clade 2.3.2 AIV have been not detected in human This is JITV Vol 26 No 3 Th. 2021:115-123 possible that the H5N1 clade 2.3.2 can be transmitted to human in Indonesia. The antiviral drugs for the prophylactic control and therapeutics of influenza infection in humans is distinguished into two classes involve the M2 inhibitors (amantadine and rimantadine) and the neuraminidase inhibitors (oseltamivir and zanamivir) (Ison 2013)

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