title: Hybrid DNA virus in Chinese patients with seronegative hepatitis discovered by deep sequencing.
abstract: Seronegative hepatitis--non-A, non-B, non-C, non-D, non-E hepatitis--is poorly characterized but strongly associated with serious complications. We collected 92 sera specimens from patients with non-A-E hepatitis in Chongqing, China between 1999 and 2007. Ten sera pools were screened by Solexa deep sequencing. We discovered a 3,780-bp contig present in all 10 pools that yielded BLASTx E scores of 7e-05-0.008 against parvoviruses. The complete sequence of the in silico-assembled 3,780-bp contig was confirmed by gene amplification of overlapping regions over almost the entire genome, and the virus was provisionally designated NIH-CQV. Further analysis revealed that the contig was composed of two major ORFs. By protein BLAST, ORF1 and ORF2 were most homologous to the replication-associated protein of bat circovirus and the capsid protein of porcine parvovirus, respectively. Phylogenetic analysis indicated that NIH-CQV is located at the interface of Parvoviridae and Circoviridae. Prevalence of NIH-CQV in patients was determined by quantitative PCR. Sixty-three of 90 patient samples (70%) were positive, but all those from 45 healthy controls were negative. Average virus titer in the patient specimens was 1.05 e4 copies/µL. Specific antibodies against NIH-CQV were sought by immunoblotting. Eighty-four percent of patients were positive for IgG, and 31% were positive for IgM; in contrast, 78% of healthy controls were positive for IgG, but all were negative for IgM. Although more work is needed to determine the etiologic role of NIH-CQV in human disease, our data indicate that a parvovirus-like virus is highly prevalent in a cohort of patients with non-A-E hepatitis.
authors.affiliation: Hematology Branch, National Heart, Lung, and Blood Institute, Bethesda, MD 20892, USA.
authors.forename: Baoyan
authors.initials: B
authors.lastname: Xu
authors.forename: Ning
authors.initials: N
authors.lastname: Zhi
authors.forename: Gangqing
authors.initials: G
authors.lastname: Hu
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Wan
authors.forename: Xiaobin
authors.initials: X
authors.lastname: Zheng
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Liu
authors.forename: Susan
authors.initials: S
authors.lastname: Wong
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kajigaya
authors.forename: Keji
authors.initials: K
authors.lastname: Zhao
authors.forename: Qing
authors.initials: Q
authors.lastname: Mao
authors.forename: Neal S
authors.initials: NS
authors.lastname: Young
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000741:Anemia, Aplastic / Q000453:epidemiology* / Q000821:virology*; D044466:Asian People / Q000706:statistics & numerical data*; D002648:Child; D002681:China / Q000453:epidemiology; D018136:Circoviridae / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D019143:Evolution, Molecular; D005260:Female; D006508:Hepatitis Antibodies / Q000097:blood; D006525:Hepatitis, Viral, Human / Q000453:epidemiology* / Q000821:virology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010321:Parvoviridae / Q000235:genetics; D010802:Phylogeny; D015995:Prevalence; D012307:Risk Factors; D016036:Seroepidemiologic Studies; D055815:Young Adult
chemical_list: D004279:DNA, Viral; D006508:Hepatitis Antibodies
keywords: Solexa sequencing; hepatitis-associated aplastic anemia; liver failure; virological diagnosis