Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing

Clinical studies indicate the etiology of liver disease to be unknown in 5% to 30% of patients. A long-standing hypothesis is the existence of unknown viruses beyond hepatitis A through E virus. We conducted serum virome sequencing in nine patients with cryptogenic liver disease and identified eight...

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Main Authors: Xiaoan Zhang, Ida X. Fan, Yanjuan Xu, Jody Rule, Long Ping Victor Tse, Mahmoud Reza Pourkarim, William M. Lee, Adrian M. Di Bisceglie, Xiaofeng Fan
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/17/6/812
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author Xiaoan Zhang
Ida X. Fan
Yanjuan Xu
Jody Rule
Long Ping Victor Tse
Mahmoud Reza Pourkarim
William M. Lee
Adrian M. Di Bisceglie
Xiaofeng Fan
author_facet Xiaoan Zhang
Ida X. Fan
Yanjuan Xu
Jody Rule
Long Ping Victor Tse
Mahmoud Reza Pourkarim
William M. Lee
Adrian M. Di Bisceglie
Xiaofeng Fan
author_sort Xiaoan Zhang
collection DOAJ
description Clinical studies indicate the etiology of liver disease to be unknown in 5% to 30% of patients. A long-standing hypothesis is the existence of unknown viruses beyond hepatitis A through E virus. We conducted serum virome sequencing in nine patients with cryptogenic liver disease and identified eight contigs that could not be annotated. One was determined to be a contaminant, while two of seven contigs from an individual (Patient 3) were validated by reverse transcription and polymerase chain reaction (RT-PCR) and Sanger sequencing. The possibility of contamination was completely excluded through PCR, with templates extracted using different methods from samples taken at different time points. One of the contigs, Seq260, was characterized as negative-sense single-stranded DNA via enzymatic digestion and genome walking. Digital-droplet PCR revealed the copy number of Seq260 to be low: 343 copies/mL. Seq260-based nested PCR screening was negative in 200 blood donors and 225 patients with liver disease with/without known etiologies. None of the seven contigs from Patient 3 was mapped onto 118,713 viral metagenomic data. Conclusively, we discovered seven unknown contigs from a patient with cryptogenic liver cirrhosis. These sequences are likely from a novel human virus with a negative-sense, linear single-stranded DNA genome.
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spelling doaj-art-4644f2e369e34bc29a87ae6b27728efb2025-08-20T03:32:32ZengMDPI AGViruses1999-49152025-06-0117681210.3390/v17060812Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome SequencingXiaoan Zhang0Ida X. Fan1Yanjuan Xu2Jody Rule3Long Ping Victor Tse4Mahmoud Reza Pourkarim5William M. Lee6Adrian M. Di Bisceglie7Xiaofeng Fan8Division of Gastroenterology & Hepatology, Department of Internal Medicine, Saint Louis University School of Medicine, St. Louis, MO 63104, USADepartment of Microbiology & Immunology, Saint Louis University School of Medicine, St. Louis, MO 63104, USADivision of Gastroenterology & Hepatology, Department of Internal Medicine, Saint Louis University School of Medicine, St. Louis, MO 63104, USADivision of Digestive and Liver Diseases, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Microbiology & Immunology, Saint Louis University School of Medicine, St. Louis, MO 63104, USALaboratory for Clinical and Epidemiological Virology, Rega Institute, Department of Microbiology, Immunology and Transplantation, KU Leuven, 3000 Leuven, BelgiumDivision of Digestive and Liver Diseases, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADivision of Gastroenterology & Hepatology, Department of Internal Medicine, Saint Louis University School of Medicine, St. Louis, MO 63104, USADivision of Gastroenterology & Hepatology, Department of Internal Medicine, Saint Louis University School of Medicine, St. Louis, MO 63104, USAClinical studies indicate the etiology of liver disease to be unknown in 5% to 30% of patients. A long-standing hypothesis is the existence of unknown viruses beyond hepatitis A through E virus. We conducted serum virome sequencing in nine patients with cryptogenic liver disease and identified eight contigs that could not be annotated. One was determined to be a contaminant, while two of seven contigs from an individual (Patient 3) were validated by reverse transcription and polymerase chain reaction (RT-PCR) and Sanger sequencing. The possibility of contamination was completely excluded through PCR, with templates extracted using different methods from samples taken at different time points. One of the contigs, Seq260, was characterized as negative-sense single-stranded DNA via enzymatic digestion and genome walking. Digital-droplet PCR revealed the copy number of Seq260 to be low: 343 copies/mL. Seq260-based nested PCR screening was negative in 200 blood donors and 225 patients with liver disease with/without known etiologies. None of the seven contigs from Patient 3 was mapped onto 118,713 viral metagenomic data. Conclusively, we discovered seven unknown contigs from a patient with cryptogenic liver cirrhosis. These sequences are likely from a novel human virus with a negative-sense, linear single-stranded DNA genome.https://www.mdpi.com/1999-4915/17/6/812etiologyhepatitis viruscryptogenic liver diseasecryptogenic cirrhosisvirome
spellingShingle Xiaoan Zhang
Ida X. Fan
Yanjuan Xu
Jody Rule
Long Ping Victor Tse
Mahmoud Reza Pourkarim
William M. Lee
Adrian M. Di Bisceglie
Xiaofeng Fan
Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
Viruses
etiology
hepatitis virus
cryptogenic liver disease
cryptogenic cirrhosis
virome
title Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
title_full Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
title_fullStr Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
title_full_unstemmed Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
title_short Novel Viral Sequences in a Patient with Cryptogenic Liver Cirrhosis Revealed by Serum Virome Sequencing
title_sort novel viral sequences in a patient with cryptogenic liver cirrhosis revealed by serum virome sequencing
topic etiology
hepatitis virus
cryptogenic liver disease
cryptogenic cirrhosis
virome
url https://www.mdpi.com/1999-4915/17/6/812
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