Scientific Publications by FDA Staff

Clin Infect Dis 2014 Jul 1;59(1):16-23

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

Nyan DC, Ulitzky LE, Cehan N, Williamson P, Winkelmann V, Rios M, Taylor DR

Background. Hepatitis B virus (HBV) is an important blood-borne pathogen that causes hepatic inflammation, and could 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%. Conclusion. These results demonstrate that our HBV-LAMP-assay is rapid, sensitive and specific, and is capable of detecting the major HVB 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.