International Journal of Medical Sciences

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20 October 2017

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Int J Med Sci 2014; 11(1):60-64. doi:10.7150/ijms.7728

Research Paper

Ultra-Deep Sequencing Analysis of the Hepatitis A Virus 5'-Untranslated Region among Cases of the Same Outbreak from a Single Source

Shuang Wu1,*, Shingo Nakamoto2,*, Tatsuo Kanda1,*, ✉, Xia Jiang1, Masato Nakamura1, Tatsuo Miyamura1, Hiroshi Shirasawa2, Nobuyuki Sugiura3, Azusa Takahashi-Nakaguchi4, Tohru Gonoi4, Osamu Yokosuka1

1. Department of Gastroenterology and Nephrology, Chiba University, Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba (260-8677), Japan
2. Department of Molecular Virology, Chiba University, Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba (260-8677), Japan
3. Department of Gastroenterology, National Hospital Organization Chiba Medical Center, 4-1-2 Tsubakimori, Chiba (260-8606), Japan
4. Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba (260-8673), Japan.
* These authors contributed equally.

Abstract

Hepatitis A virus (HAV) is a causative agent of acute viral hepatitis for which an effective vaccine has been developed. Here we describe ultra-deep pyrosequences (UDPSs) of HAV 5'-untranslated region (5'UTR) among cases of the same outbreak, which arose from a single source, associated with a revolving sushi bar. We determined the reference sequence from HAV-derived clone from an attendant by the Sanger method. Sixteen UDPSs from this outbreak and one from another sporadic case were compared with this reference. Nucleotide errors yielded a UDPS error rate of < 1%. This study confirmed that nucleotide substitutions of this region are transition mutations in outbreak cases, that insertion was observed only in non-severe cases, and that these nucleotide substitutions were different from those of the sporadic case. Analysis of UDPSs detected low-prevalence HAV variations in 5'UTR, but no specific mutations associated with severity in these outbreak cases. To our surprise, HAV strains in this outbreak conserved HAV IRES sequence even if we performed analysis of UDPSs. UDPS analysis of HAV 5'UTR gave us no association between the disease severity of hepatitis A and HAV 5'UTR substitutions. It might be more interesting to perform ultra-deep sequencing of full length HAV genome in order to reveal possible unknown genomic determinants associated with disease severity. Further studies will be needed.

Keywords: Acute liver failure, HAV, IRES, UDPS, 5'UTR.

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Wu S, Nakamoto S, Kanda T, Jiang X, Nakamura M, Miyamura T, Shirasawa H, Sugiura N, Takahashi-Nakaguchi A, Gonoi T, Yokosuka O. Ultra-Deep Sequencing Analysis of the Hepatitis A Virus 5'-Untranslated Region among Cases of the Same Outbreak from a Single Source. Int J Med Sci 2014; 11(1):60-64. doi:10.7150/ijms.7728. Available from http://www.medsci.org/v11p0060.htm