CRISPR/Cas13a-assisted rapid and portable HBV DNA detection for low-level viremia patients. Issue 1 (31st December 2023)

• Record Type: Journal Article
• Authors: Tian, Yuan; Fan, Zihao; Xu, Ling; Cao, Yaling; Chen, Sisi; Pan, Zhenzhen; Gao, Yao; Li, Hao; Zheng, Sujun; Ma, Yingmin; Duan, Zhongping; Zhang, Xiangying; Ren, Feng
• Is Part Of: Emerging microbes & infections. Volume 12:Issue 1(2023)
• Publication Date: 2023-12-31
• Subjects: HBV DNA -- CRISPR/Cas13a -- Low-level viremia -- Recombinase-aided amplification (RAA) -- Detection; Medical microbiology -- Periodicals; Communicable diseases -- Periodicals; Infection -- Periodicals; 616.9041
• DOI: 10.1080/22221751.2023.2177088
• Languages: English
• ISSNs: 2222-1751

Abstract

ABSTRACT: Background & Aims: The WHO declared to eliminate hepatitis B virus (HBV) by 2030. However, an increasing number of patients are presenting with low-level viremia (LLV) with the widespread use of antiviral medications. The diagnostic efficiency and coverage area of HBV infection are low. Hence, this study intended to drive the HBV infection detection to effectively adaptable for any small to medium-sized laboratory or field survey. Methods: We established, optimized, and evaluated a colloidal gold test strip for detection of HBV DNA based on CRISPR/Cas13a combined with recombinase-aided amplification (RAA) technology. Furthermore, 180 HBV-infected patients (including patients with different viral loads, LLV patients and dynamic plasma samples of patients on antiviral therapy) were enrolled for clinical validation. Results: The strip detection of HBV DNA was established based on RAA-CRISPR-Cas13a technology with a sensitivity of 10 1 copies/μL and a specificity of 100%. HBV DNA gradient concentration plasmids and clinical samples were effectively identified by this approach. The positive coincidence rate for LLV patients was 87%, while the negative coincidence rate was 100%. The positive coincidence rate reached 100% in LLV patients (viral loading >100 IU/mL). The sensitivity, specificity, positive predictive agreement (PPA) and negative predictive agreement (NPA) values of dynamic plasma detection in patients on antiviral therapy were 100%, 92.15%, 93.75%, and 100%, respectively. Conclusions: We develop rapid and portable RAA-CRISPR/Cas13a-based strip of HBV DNA detection for LLV patients. This study provides a visual and faster alternative to current PCR-based diagnosis for HBV infection.