Journal Article

Rapid Detection of Hepatitis B Virus in Blood Plasma by a Specific and Sensitive Loop-Mediated Isothermal Amplification Assay

Dougbeh-Chris Nyan, Laura E. Ulitzky, Nicoleta Cehan, Phillip Williamson, Valerie Winkelman, Maria Rios and Deborah R. Taylor

in Clinical Infectious Diseases

Published on behalf of Infectious Diseases Society of America

Volume 59, issue 1, pages 16-23
Published in print July 2014 | ISSN: 1058-4838
Published online April 2014 | e-ISSN: 1537-6591 | DOI: https://dx.doi.org/10.1093/cid/ciu210
Rapid Detection of Hepatitis B Virus in Blood Plasma by a Specific and Sensitive Loop-Mediated Isothermal Amplification Assay

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Background. Hepatitis B virus (HBV) is an important blood-borne pathogen that causes hepatic inflammation and can lead to liver cirrhosis and hepatocellular carcinoma. Conventional methods of HBV detection are time consuming and require highly trained personnel and elaborate equipment. This report describes the development of a rapid, simple, specific, and sensitive loop-mediated isothermal amplification assay (LAMP) for detection of HBV genotypes A, B, C, D, E, and F in blood samples.

Methods. HBV standard plasma panels and clinical donor plasma specimens were used for the development and validation of the LAMP assay. Amplification was performed at 60°C for 60 minutes using extracted DNA or heat-treated plasma specimens without DNA extraction. The assay was evaluated for its ability to detect various HBV genotypes and for its sensitivity, specificity, and time-point of detection.

Results. The LAMP assay detected HBV genotypes A–F and demonstrated a sensitivity of 10–100 IU per reaction of HBV DNA. The assay also detected 69 of 75 (92%) HBV-positive donor plasma specimens tested and demonstrated a specificity of 100%.

Conclusions. These results demonstrate that our HBV-LAMP assay is rapid, sensitive and specific, and capable of detecting the major HBV genotypes. This assay could be used in clinical point-of-care settings, mainly in endemic and resource-limited environments for HBV diagnostics, donor screening, epidemiological studies, and therapeutic monitoring of patients undergoing antiviral treatment.

Keywords: hepatitis B virus; loop-mediated isothermal amplification; nucleotide; rapid detection; international units per reaction

Journal Article.  4558 words.  Illustrated.

Subjects: Infectious Diseases ; Immunology ; Public Health and Epidemiology ; Microbiology

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