Infection and Drug Resistance (Mar 2021)

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

  • Chen X,
  • Zhou Q,
  • Dong S,
  • Wang S,
  • Liu R,
  • Wu X,
  • Li S

Journal volume & issue
Vol. Volume 14
pp. 1219 – 1229

Abstract

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Xu Chen,1,2,* Qingxue Zhou,3,* Shilei Dong,4 Shuoshi Wang,2 Rui Liu,2 Xueli Wu,2 Shijun Li5 1The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, People’s Republic of China; 2Central Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, People’s Republic of China; 3Clinical Laboratory, Hangzhou Women’s Hospital, Hangzhou, Zhejiang, 310008, People’s Republic of China; 4Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, Zhejiang, 310013, People’s Republic of China; 5Laboratory of Bacterial Infectious Disease of Experimental Centre, Guizhou Provincial Centre for Disease Control and Prevention, Guiyang, Guizhou, 550004, People’s Republic of China*These authors contributed equally to this workCorrespondence: Shijun LiGuizhou Provincial Centre for Disease Control and Prevention, 115 Bageyan Road, Guiyang, Guizhou, 550004, People’s Republic of ChinaTel/Fax +86 851 86824290Email [email protected] ChenThe Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, 83 Feishan Road, Guiyang, Guizhou, 550003, People’s Republic of ChinaTel/Fax +86 851 85514432Email [email protected]: 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.Keywords: hepatitis B virus, HBV, multiple cross displacement amplification, MCDA, lateral flow biosensor, LFB, rapid detection, molecular detection

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