Detection of Hepatitis B Virus by Loop Mediated Isothermal Amplification (LAMP)

Hepatitis B virus (HBV) belongs to hepadnaviridea family which is one of the main factors of hepatocellular carcinoma and liver diseases. The HBV detection methods (serological and molecular) have their own limitations, so the using ability of them in all diagnosis centres is not possible. In this study it has been tried to apply new novel LAMP technique on serum samples by using specific primer designed for HBs region. 104 HBV quantities sera by COBAS Amplicor Monitor kit were supplied. DNA was extracted by DNP kit and then PCR primers were selected and 6 specific primers were designed for LAMP. PCR and LAMP sensitivity and specificity were determined. PCR product by electrophoresis and LAMP product by adding 0.1% SYBR Green were identified. Among 104 quantitative serum, only 95 cases were PCR positive but 101 cases were LAMP positive.9 cases were PCR negative among these, 6 cases were reported as LAMP positive. In 3 cases LAMP and PCR were both negative. Application compression of PCR and LAMP technique on patient's samples with definite virus particles showed that the LAMP technique had more specificity and sensitivity.