Design and Evaluation of real-time PCR for Molecular Diagnosis of Hepatitis B

The hepatitis B virus (HBV) was discovered in 1965. Approximately 350 million people are infected with HBV. The virus has a worldwide spread and one of the most important complications of infection is chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Hepatitis can be detected by biochemical tests that determine liver function.

This study was qualitative and 30 patients with suspected hepatitis B referring to Massoud Lab in 2019 were selected. Blood samples were tested for HBsAg positive by multiplex RT-PCR with specific probes and primers as well as β-Actin gene as quality control. Then the obtained results were compared with the results of commercial tests.

From 30 samples, 19 samples were HBsAg positive and 11 samples were HBsAg negative. Positive samples were re-examined to confirm the positive results. To compare and evaluate the quality of the results of the designed test, all of these 30 samples were analyzed using another common laboratory kit.

The high sensitivity, good reproducibility, and low cost make this innovative quantitative HBV real-time PCR assay especially well-suited for application to large clinical surveys.