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Validation and implementation of hepatitis b viral load detection with quantitative real-time PCR

Author: 
Shiva V Murarka, Khushbu R Patel, Parth S Shah, Nidhi D Shah, Bhavini S Shah and Mandava V Rao
Subject Area: 
Health Sciences
Abstract: 

Hepatitis B viral (HBV) load monitoring is mainly important for the patients suffering from liver related diseases. Several techniques have been established to identify the viral load in the patients such as serological tests and various types of PCR. In this cohort we describe the validation and implementation of cost-effective quantitative real-time PCR for Hepatitis B viral load detection. The use of Quantstudio 3D digital PCR for the preparation of in-house standards also makes the technology more cost-effective and sensitive. The real-time HBV assay was validated for its linearity, sensitivity, precision, reproducibility while at the same time we also compared with the digital PCR (dPCR) for its accuracy. The sensitivity of digital PCR (dPCR) is better than RT-PCR due to its absolute quantification. RT-PCR is used in all laboratories with Taqman chemistry which is almost competent with dPCR, though the latter is better in a way that no standards or controls are required for quantification.

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