Multiple Cross Displacement Amplification Linked with Nanoparticles-Based Lateral Flow Biosensor in Screening of Hepatitis B Virus in Clinical Application

Abstract

Background Hepatitis B virus (HBV) is a common pathogen that predominantly causes severe liver disease, and remains one of a huge challenge worldwide, especially in many resource-constrained areas. Developing a low-cost, sensitive, specific, and rapid approach for screening HBV is critical for its treatment and prevention. In the current study, a novel molecular detection approach, multiple cross displacement amplification (MCDA) coupled with polymer nanoparticle-based lateral flow biosensor (MCDA-LFB), was applied for detection of HBV in blood samples. Methods HBV standard substance and clinical donor serum samples were collected and used for the establishment and confirmation of the HBV-MCDA-LFB assay. A set of 10 MCDA primers was designed according to HBV-specific gene S . The HBV-MCDA-LFB assay conditions, including genomic template concentration, MCDA reaction temperature and time were optimized. The sensitivity and specificity of the HBV-MCDA -LFB assay were evaluated in this report. The HBV-MCDA-LFB assay was applied to detect the HBV agent from clinical samples. Results The HBV-MCDA primers based on the S gene were valid for establishment of MCDA assay. The HBV-MCDA reaction with optimized conditions could be carried out at a constant temperature 64°C for 35 min. The whole process, including sample preparation (5 min), genomic template extraction (~30 min), MCDA amplification (35 min), and LFB reading (~2 min), could be completed within 80 min. The sensitivity of this assay was 5 IU per reaction. The specificity was 100% for HBV-MCDA-LFB assay. Conclusion These results confirmed that the HBV-MCDA-LFB is a low-cost, sensitive, specific, simple, and rapid method for detecting HBV agents. This technique has great potential to develop a point-of-care testing (POCT) method in clinical practice, especially in endemic and resource-constrained regions.

Applications of laboratory findings in the prevention, diagnosis, treatment, and monitoring of COVID-19

“…Lateral-flow assay (LFA) used for the rapid SARS-CoV-2 serological detection. Apart from ELISA and CLIA/ECLIA, the lateral-flow assay (LFA) is a paper-based platform for the rapid detection and quantification assay that is wildly used in the antibody and antigen detection of pathogens (like the influenza virus, 143 HBV, 144 and HCV 145 ). Generally, the platform is composed of three parts: sample pad, conjugate pad, and detection pad.…”

Highly sensitive and rapid identification of coxsackievirus A16 based on reverse transcription multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor assay

“…Multiple cross displacement amplification (MCDA), a novel, inexpensive, quick, easy, and effective isothermal amplification approach identical to LAMP that has been widely employed in the identification of various pathogens, such as bacteria, viruses, and fungi, has been developed to address the limitations of traditional identification ( Li et al, 2019 ; Wang et al, 2021 ; Chen et al, 2021b ). To accomplish targeted amplification using the MCDA approach, 10 primers that bind to 10 different regions of the target sequence must be designed.…”

A Nanoparticle-Based Biosensor Combined With Multiple Cross Displacement Amplification for the Rapid and Visual Diagnosis of Neisseria gonorrhoeae in Clinical Application

“…The strategy has potential applications as a point-of-care (POC) assay due to its rapidity, simplicity, and easy-to-use operations. Critically, it has been used to detect several pathogens such as SARS-CoV-2, hepatitis B virus, and Streptococcus agalactiae ( Cheng et al, 2020 ; Li et al, 2020 ; Chen et al, 2021 ). The gold nanoparticle-based lateral flow biosensor (LFB) is a paper-based platform, which facilitates a low-cost rapid diagnosis; it is robust, visual, sensitive, specific, and has a low limit of detection (LoD) ( Zeng et al, 2013 ; Urusov et al, 2019 ).…”