A Novel Colorimetric PCR-Based Biosensor for Detection and Quantification of Hepatitis B Virus

Abstract

We have established an operationconvenient and cost-effective way for detection HBV. This method has shown good performance with a broad range of linearity and high sensitivity. The results of detection is reported with macroscopic colorimetric signals. The strategy takes the key advantage of the TaqMan technology and use inexpensive DNA sensor. This approach can detect HBV DNA qualitatively or quantificationally. Hepatitis B virus (HBV) can cause viral infection that attacks the liver and it is a major global health problem that put people at a high risk of death from cirrhosis of the liver and liver cancer. HBV has infected one third of the worldwide population, and 350 million people suffer from chronic HBV infection. For these reasons, development of an accurate, sensitive and expedient detection method for diagnosing, monitoring and assessing therapeutic response of HBV is very necessary and urgent for public health and disease control. Here we report a new strategy for detection of viral load quantitation of HBV based on colorimetric polymerase chain reaction (PCR) with DNAzyme-containing probe. The special DNAzyme adopting a G-quadruplex structure exhibited peroxidase-like activity in the presence of hemin to report colorimetric signal. This method has shown a broad range of linearity and high sensitivity. This study builds important foundation to achieve the specific and accurate detection level of HBV DNA with a low-cost and effective method in helping diagnosing, preventing and protecting human health form HBV generally all over the world and especially in developing countries.

Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor

“…Previous reports have employed a similar strategy to detect both human and plant viruses (Yang et al, 2017;Kim et al, 2018;Wang et al, 2019), and a recent report has suggested the potential use of DNAzyme sensors for SARS-CoV-2 detection (Xi et al, 2020). We here demonstrate the utility of this approach for SARS-CoV-2.…”

“…We followed a previous strategy of tagging the 5 ′ end of the primers with sequences complementary to the G-quadruplex which forms a G-quadruplex only upon amplification of the PCR product (Bhadra et al, 2014). A similar strategy has been employed for the detection of viruses such as hepatitis B virus and to detect the HIV-1 gag gene by other studies previously (Yang et al, 2017;Kim et al, 2018). We employed this approach to detect SARS-CoV-2 RNA by conventional PCR.…”

Visual Detection of Cucumber Green Mottle Mosaic Virus Based on Terminal Deoxynucleotidyl Transferase Coupled with DNAzymes Amplification

“…Visual nucleic acid detection has attracted much attention due to its simplicity, high sensitivity, and visual results [22,23,24,25]. For example, colorimetric detection has been reported for some animal viruses, including H1N1 influenza virus [26,27,28,29], H5N1 influenza virus [30], hepatitis B virus [31,32], Zika virus [33], and Rift Valley Fever Virus [34]. A colorimetric detection method was reported for CGMMV [35] based on unmodified gold nanoparticles, with a limited detection of 30 pg/μL of CGMMV RNA.…”