Development of a sensitive novel diagnostic kit for the highly pathogenic avian influenza A (H5N1) virus.

Yasuko Tsunetsugu-Yokota,Mie Kobayashi-Ishihara,Tsutomu Kageyama,Kazuo Ohnishi,Shuhei Misawa,Ikuyo Takayama,Kengo Nishimura,Giang T Dang,Long T Nguyen,Hang Lk Nguyen,Masato Tashiro,Shigeyuki Itamura,Hitoshi Takahashi,Mai Tq Le

doi:10.1186/1471-2334-14-362

Abstract

BackgroundSporadic emergence of the highly pathogenic avian influenza (HPAI) H5N1 virus infection in humans is a serious concern because of the potential for a pandemic. Conventional or quantitative RT-PCR is the standard laboratory test to detect viral influenza infections. However, this technology requires well-equipped laboratories and highly trained personnel. A rapid, sensitive, and specific alternative screening method is needed.MethodsBy a luminescence-linked enzyme immunoassay, we have developed a H5N1 HPAI virus detection kit using anti-H5 hemagglutinin monoclonal antibodies in combination with the detection of a universal NP antigen of the type A influenza virus. The process takes 15 minutes by use of the fully automated luminescence analyzer, POCube.ResutlsWe tested this H5/A kit using 19 clinical specimens from 13 patients stored in Vietnam who were infected with clade 1.1 or clade 2.3.4 H5N1 HPAI virus. Approximately 80% of clinical specimens were H5-positive using the POCube system, whereas only 10% of the H5-positive samples were detected as influenza A-positive by an immunochromatography-based rapid diagnostic kit.ConclusionsThis novel H5/A kit using POCube is served as a rapid and sensitive screening test for H5N1 HPAI virus infection in humans.

Highlights

Sporadic emergence of the highly pathogenic avian influenza (HPAI) H5N1 virus infection in humans is a serious concern because of the potential for a pandemic
We evaluated this novel H5/A diagnostic kit using clinical specimens infected with the H5N1 HPAI virus in Vietnam and demonstrated the sensitive dual detection of H5 HA and type A nucleoprotein (NP) antigens for the first time
Despite of a limited number of available H5N1 clinical specimens, our results strongly suggest that this diagnostic test is a useful tool in the rapid and reliable identification of H5N1 HPAI virus infections

Summary

Introduction

Sporadic emergence of the highly pathogenic avian influenza (HPAI) H5N1 virus infection in humans is a serious concern because of the potential for a pandemic. Conventional or quantitative RT-PCR is the standard laboratory test to detect viral influenza infections. This technology requires well-equipped laboratories and highly trained personnel. The high variability of influenza A viruses is driven by frequent mutations in genomic RNA (drift) and by genetic reassortment among avian, porcine, and human strains [4]. This has hampered the development of a universal cross-protective flu vaccine

Methods

Results

Conclusion