Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform
The recent discovery of collateral cleavage activity of class-II clustered regularly interspaced short palindromic repeats–CRISPR-associated protein (CRISPR-Cas) makes CRISPR-based diagnosis a potential high-accuracy nucleic acid detection method. Colloidal gold-based lateral flow immunochromatograp...
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doaj-30061bed531f4f73bfeac3c0a21c992b2021-07-07T06:15:59ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-07-011210.3389/fmicb.2021.700016700016Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA PlatformYang YouPingping ZhangGengshan WuYafang TanYong ZhaoShiyang CaoYajun SongRuifu YangZongmin DuThe recent discovery of collateral cleavage activity of class-II clustered regularly interspaced short palindromic repeats–CRISPR-associated protein (CRISPR-Cas) makes CRISPR-based diagnosis a potential high-accuracy nucleic acid detection method. Colloidal gold-based lateral flow immunochromatographic assay (LFA), which has been combined with CRISPR/Cas-based nucleic detection, usually associates with drawbacks of relative high background and the subjectivity in naked-eye read-out of the results. Here, we developed a novel system composed of Cas12a-based nucleic acid detection and up-converting phosphor technology (UPT)-based LFA (UPT–LFA), termed Cas12a-UPTLFA. We further demonstrated the utility of this platform in highly sensitive and specific detection of Yersinia pestis, the causative agent of the deadly plague. Due to high infectivity and mortality, as well as the potential to be misused as bioterrorism agent, a culture-free, ultrasensitive, specific, and rapid detection method for Y. pestis has long been desired. By incorporating isothermal recombinase polymerase amplification, the Cas12a-UPTLFA we established can successfully detect genomic DNA of Y. pestis as low as 3 attomolar (aM) and exhibited high sensitivity (93.75%) and specificity (90.63%) for detection of spiked blood samples with a detection limit of 102 colony-forming unit per 100 μl of mouse blood. With a portable biosensor, Cas12a-UPTLFA assay can be operated easily by non-professional personnel. Taken together, we have developed a novel Cas12a-UPTLFA platform for rapid detection of Y. pestis with high sensitivity and specificity, which is portable, not expensive, and easy to operate as a point-of-care method. This detection system can easily be extended to detect other pathogens and holds great promise for on-site detection of emerging infectious pathogens.https://www.frontiersin.org/articles/10.3389/fmicb.2021.700016/fulllateral flow immunochromatographic assayup-converting phosphor technologynucleic acid detectionYersinia pestisCas12a |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yang You Pingping Zhang Gengshan Wu Yafang Tan Yong Zhao Shiyang Cao Yajun Song Ruifu Yang Zongmin Du |
spellingShingle |
Yang You Pingping Zhang Gengshan Wu Yafang Tan Yong Zhao Shiyang Cao Yajun Song Ruifu Yang Zongmin Du Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform Frontiers in Microbiology lateral flow immunochromatographic assay up-converting phosphor technology nucleic acid detection Yersinia pestis Cas12a |
author_facet |
Yang You Pingping Zhang Gengshan Wu Yafang Tan Yong Zhao Shiyang Cao Yajun Song Ruifu Yang Zongmin Du |
author_sort |
Yang You |
title |
Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform |
title_short |
Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform |
title_full |
Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform |
title_fullStr |
Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform |
title_full_unstemmed |
Highly Specific and Sensitive Detection of Yersinia pestis by Portable Cas12a-UPTLFA Platform |
title_sort |
highly specific and sensitive detection of yersinia pestis by portable cas12a-uptlfa platform |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2021-07-01 |
description |
The recent discovery of collateral cleavage activity of class-II clustered regularly interspaced short palindromic repeats–CRISPR-associated protein (CRISPR-Cas) makes CRISPR-based diagnosis a potential high-accuracy nucleic acid detection method. Colloidal gold-based lateral flow immunochromatographic assay (LFA), which has been combined with CRISPR/Cas-based nucleic detection, usually associates with drawbacks of relative high background and the subjectivity in naked-eye read-out of the results. Here, we developed a novel system composed of Cas12a-based nucleic acid detection and up-converting phosphor technology (UPT)-based LFA (UPT–LFA), termed Cas12a-UPTLFA. We further demonstrated the utility of this platform in highly sensitive and specific detection of Yersinia pestis, the causative agent of the deadly plague. Due to high infectivity and mortality, as well as the potential to be misused as bioterrorism agent, a culture-free, ultrasensitive, specific, and rapid detection method for Y. pestis has long been desired. By incorporating isothermal recombinase polymerase amplification, the Cas12a-UPTLFA we established can successfully detect genomic DNA of Y. pestis as low as 3 attomolar (aM) and exhibited high sensitivity (93.75%) and specificity (90.63%) for detection of spiked blood samples with a detection limit of 102 colony-forming unit per 100 μl of mouse blood. With a portable biosensor, Cas12a-UPTLFA assay can be operated easily by non-professional personnel. Taken together, we have developed a novel Cas12a-UPTLFA platform for rapid detection of Y. pestis with high sensitivity and specificity, which is portable, not expensive, and easy to operate as a point-of-care method. This detection system can easily be extended to detect other pathogens and holds great promise for on-site detection of emerging infectious pathogens. |
topic |
lateral flow immunochromatographic assay up-converting phosphor technology nucleic acid detection Yersinia pestis Cas12a |
url |
https://www.frontiersin.org/articles/10.3389/fmicb.2021.700016/full |
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