Loop mediated isothermal amplification (LAMP) for rapid detection of HBV in Iran

Abstract

Hepatitis B virus (HBV) is one of the most important factors for hepatocellular carcinoma and a liver disease that seems 350 - 400 million persons are infected with all over the world. Loop mediated isothermal amplification (LAMP) is a novel nucleic acid amplification technique with high specificity and sensitivity which has been done under isothermal condition. In this study we tried to apply LAMP technique for rapid, accurate, and cost-effective diagnosis of HBV in patient's serum samples. 104 HBV quantities sera were supplied. Six LAMP specific primers were designed for HBs region of HBV. LAMP and PCR reactions were optimized. At the end of the LAMP reaction, SYBR Green was used for identifying negative and positive results. The PCR sensitivity up to 40 particles was observed and the LAMP sensitivity test was verified up to 4 particles. Among the 104 quantities sera, 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. In comparison, between LAMP and PCR, the LAMP technique in spite of its simplicity, high sensitivity and specificity, could be an appropriate replacement for PCR.                                                    Key word: Hepatitis B, PCR, LAMP.