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A pilot study of serum microRNA signatures as a novel biomarker for occult hepatitis B virus infection

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Abstract

The implementation of hepatitis B surface antigen (HBsAg) screening tests has significantly enhanced blood transfusion safety. However, the transmission of HBsAg-negative blood components can still occur in the acute phase of infection during the seronegative window period or during chronic stages of infection such as occult hepatitis virus B infection (OBI). OBI, characterized by the presence of HBV infection without detectable HBsAg, is capable to elude the routine detection with HBV serologic markers and harbor a potential risk of HBV transmission through blood transfusion or organ transplantation. Here, we test the hypothesis that OBI patients have a differentially expressed profile of microRNA (miRNA) in serum, and this unique serum miRNA signature can serve as a biomarker to detect OBI. Employing TaqMan probe-based quantitative reverse transcription polymerase chain reaction (qRT-PCR), we assessed the expression level of miRNAs in serum samples. To control for miRNA quantitation, we added an exogenous plant miRNA, MIR156a, into the samples before RNA extraction and used it as an internal control. After screening 13 previously identified HBV-specific serum miRNAs, we obtained four miRNAs, let-7c, miR-23b, miR-122, and miR-150, which are differentially expressed in OBI sera compared to healthy control sera. This 4-serum miRNA signature shows a high level of accuracy in discriminating both OBI (AUC = 0.999) and HBV (AUC = 0.989) cases from the non-infected controls. Cluster analysis also demonstrates that this 4-miRNA signature can clearly separate OBI patients from the control group. Our results demonstrate for the first time that a profile of serum miRNAs can serve as a sensitive and accurate biomarker for OBI detection.

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Fig. 1

Abbreviations

HBV:

Hepatitis B virus

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

OBI:

Occult HBV infection

HBsAg:

Hepatitis B surface antigen

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

AUC:

Area under curve

ROC:

Receiver operating characteristic

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Acknowledgments

The authors thank Dr. Xi Chen and Ms. Qiang Zhang for excellent assistance in sample collection and preparation. This work was supported by grants from the National Natural Science Foundation of China [(no. 30988003, 30871019, 31000323, and 90608010), the National Basic Research Program of China (973 program no. 2011CB504803)].

Conflict of interest

The authors have no conflict of interest to declare.

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Authors and Affiliations

  1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 22 Hankou Road, Nanjing, 210093, China

    Yuanyuan Chen, Limin Li, Nan Wang, Chen-Yu Zhang & Ke Zen

  2. Clinical Laboratory, Nanjing Second Hospital, Nanjing, 210003, China

    Zhenxian Zhou

Authors
  1. Yuanyuan Chen
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  2. Limin Li
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  3. Zhenxian Zhou
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  4. Nan Wang
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  5. Chen-Yu Zhang
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Corresponding authors

Correspondence to Chen-Yu Zhang or Ke Zen.

Additional information

Yuanyuan Chen, Limin Li, Zhenxian Zhou contributed equally to this work.

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Chen, Y., Li, L., Zhou, Z. et al. A pilot study of serum microRNA signatures as a novel biomarker for occult hepatitis B virus infection. Med Microbiol Immunol 201, 389–395 (2012). https://doi.org/10.1007/s00430-011-0223-0

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  • DOI: https://doi.org/10.1007/s00430-011-0223-0

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