CRISPR/Cas12-Based Ultra-Sensitive and Specific Point-of-Care Detection of HBV.
CRISPR/Cas12-Based Ultra-Sensitive and Specific Point-of-Care Detection of HBV.
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DOI：
10.3390/ijms22094842
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发表时间：
2021-05-03
影响因子：
5.6
通讯作者：
Duan G
中科院分区：
文献类型：
--
作者：
Ding R;Long J;Yuan M;Zheng X;Shen Y;Jin Y;Yang H;Li H;Chen S;Duan G
B型肝炎仍然是一个重大的全球公共卫生挑战，在医疗条件差的西太平洋和非洲地区发病率特别高。尽管临床上用于HBV qPCR检测的技术已经成熟，但对床旁检测的研究尚未取得实质性进展。开发快速、准确的床旁检测对于医疗条件差的农村地区预防和控制B型肝炎至关重要。CRISPR/Cas系统在核酸检测中的发展允许病原体即时检测。在这里，我们开发了一种基于LAMP-Cas 12 a的快速准确的HBV即时检测方法。它创新地解决了10分钟内即时检测的问题，特别是样本核酸提取的问题。基于LAMP-Cas 12 a，通过荧光读数和侧向流测试条呈现测定结果的可视化。侧流试纸条技术可以实现肉眼可见的结果，而荧光读出可以实现实时高灵敏度检测。基于荧光读出的Cas 12 a检测方法可在13 min内实现HBV检测，检测限为1拷贝/μL，而侧流试纸条技术仅需20 min。在73份临床样本的评价中，荧光读出法和侧流试纸条法的灵敏度和特异性均为100%，检测结果与qPCR完全相当。基于LAMP-Cas 12 a的HBV检测依赖于最少的设备，以提供快速，准确的检测结果和低成本，为床旁HBV检测提供了重要的实用价值。
Hepatitis B remains a major global public health challenge, with particularly high prevalence in medically disadvantaged western Pacific and African regions. Although clinically available technologies for the qPCR detection of HBV are well established, research on point-of-care testing has not progressed substantially. The development of a rapid, accurate point-of-care test is essential for the prevention and control of hepatitis B in medically disadvantaged rural areas. The development of the CRISPR/Cas system in nucleic acid detection has allowed for pathogen point-of-care detection. Here, we developed a rapid and accurate point-of-care assay for HBV based on LAMP-Cas12a. It innovatively solves the problem of point-of-care testing in 10 min, particularly the problem of sample nucleic acid extraction. Based on LAMP-Cas12a, visualization of the assay results is presented by both a fluorescent readout and by lateral flow test strips. The lateral flow test strip technology can achieve results visible to the naked eye, while fluorescence readout can achieve real-time high-sensitivity detection. The fluorescent readout-based Cas12a assay can achieve HBV detection with a limit of detection of 1 copy/μL within 13 min, while the lateral flow test strip technique only takes 20 min. In the evaluation of 73 clinical samples, the sensitivity and specificity of both the fluorescence readout and lateral flow test strip method were 100%, and the results of the assay were fully comparable to qPCR. The LAMP-Cas12a-based HBV assay relies on minimal equipment to provide rapid, accurate test results and low costs, providing significant practical value for point-of-care HBV detection.
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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
DOI：
10.1016/s2468-1253(20)30307-1
发表时间：
2021-03
期刊：
The lancet. Gastroenterology & hepatology
影响因子：
--
作者：
Tan M;Bhadoria AS;Cui F;Tan A;Van Holten J;Easterbrook P;Ford N;Han Q;Lu Y;Bulterys M;Hutin Y
通讯作者：
Hutin Y
DOI：
10.1126/science.aav4294
发表时间：
2018-11-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Harrington LB;Burstein D;Chen JS;Paez-Espino D;Ma E;Witte IP;Cofsky JC;Kyrpides NC;Banfield JF;Doudna JA
通讯作者：
Doudna JA
CRISPR-Cas12a辅助核酸检测
影响因子：
33.5
作者：
Li SY;Cheng QX;Wang JM;Li XY;Zhang ZL;Gao S;Cao RB;Zhao GP;Wang J
通讯作者：
Wang J
影响因子：
168.9
作者：
Schweitzer, Aparna;Horn, Johannes;Ott, Joerdis J.
通讯作者：
Ott, Joerdis J.