CRISPR/Cas13a-assisted rapid and portable HBV DNA detection for low-level viremia patients.
CRISPR/Cas13a-assisted rapid and portable HBV DNA detection for low-level viremia patients.
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DOI：
10.1080/22221751.2023.2177088
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发表时间：
2023-12
影响因子：
13.2
通讯作者：
中科院分区：
文献类型：
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作者：
世界卫生组织宣布到2030年消除乙肝病毒。然而，随着抗病毒药物的广泛使用，越来越多的患者出现了低水平病毒血症(LLV)。乙肝病毒感染的诊断效率低，覆盖面低。因此，本研究旨在推动乙肝病毒感染检测有效地适用于任何中小型实验室或现场调查。我们建立、优化和评价了基于CRISPR/Cas13a和重组酶辅助扩增(RAA)技术的检测HBVDNA的胶体金试纸条。此外，180例乙肝病毒感染患者(包括不同病毒载量的患者、LLV患者和抗病毒治疗患者的动态血浆样本)进入临床验证。建立了基于RAA-CRISPR-CAS13a技术的HBVDNA纸条检测方法，其灵敏度为101Copes/μ，特异度为100%。该方法能有效地鉴定HBVDNA梯度浓度质粒和临床标本。LLV阳性符合率为87%，阴性符合率为100%。LLV患者(病毒载量100IU/mL)的阳性符合率达100%。动态血浆检测抗病毒治疗的敏感性、特异性、阳性预测值和阴性预测值分别为100%、92.15%、93.75%和100%。我们开发了基于RAA-CRISPR/CA13a的LLV患者HBVDNA快速检测纸条。这项研究为目前基于聚合酶链式反应的乙肝病毒感染诊断提供了一个直观和快速的替代方法。
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. 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. The strip detection of HBV DNA was established based on RAA-CRISPR-Cas13a technology with a sensitivity of 101 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. 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.
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DOI：
10.1126/science.abb1400
发表时间：
2020-07-17
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Pausch P;Al-Shayeb B;Bisom-Rapp E;Tsuchida CA;Li Z;Cress BF;Knott GJ;Jacobsen SE;Banfield JF;Doudna JA
通讯作者：
Doudna JA
DOI：
10.1126/science.aas8836
发表时间：
2018-04-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Myhrvold C;Freije CA;Gootenberg JS;Abudayyeh OO;Metsky HC;Durbin AF;Kellner MJ;Tan AL;Paul LM;Parham LA;Garcia KF;Barnes KG;Chak B;Mondini A;Nogueira ML;Isern S;Michael SF;Lorenzana I;Yozwiak NL;MacInnis BL;Bosch I;Gehrke L;Zhang F;Sabeti PC
通讯作者：
Sabeti PC
影响因子：
5.6
作者：
Ding R;Long J;Yuan M;Zheng X;Shen Y;Jin Y;Yang H;Li H;Chen S;Duan G
通讯作者：
Duan G
影响因子：
8.8
作者：
Bonanzinga, Sara;Onelia, Francesco;Chevaliez, Stephane
通讯作者：
Chevaliez, Stephane
影响因子：
4.3
作者：
Liu, Yue-Ping;Yao, Chun-Yan
通讯作者：
Yao, Chun-Yan