{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T14:46:43Z","timestamp":1790520403787,"version":"4.1.0"},"reference-count":24,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2022,8,1]],"date-time":"2022-08-01T00:00:00Z","timestamp":1659312000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100006593","name":"Board of Research in Nuclear Sciences","doi-asserted-by":"publisher","award":["BRNS\/37080"],"award-info":[{"award-number":["BRNS\/37080"]}],"id":[{"id":"10.13039\/501100006593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Aim<\/jats:title>\n                  <jats:p>The current scenario of COVID-19 pandemic has presented an almost insurmountable challenge even for the most sophisticated hospitals equipped with modern biomedical technology. There is an urgency to develop simple, fast and highly accurate methods for the rapid identification and isolation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected patients. To address the ongoing challenge, the present study offers a CLEVER assay (CRISPR-Cas integrated RT-LAMP Easy, Visual and Extraction-free RNA) which will allow RNA extraction-free method to visually diagnose COVID-19. RNA extraction is a major hurdle in preventing rapid and large-scale screening of samples particularly in low-resource regions because of the logistics and costs involved.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Method and Result<\/jats:title>\n                  <jats:p>Herein, the visual SARS-CoV-2 detection method consists of RNA extraction-free method directly utilizing the patient's nasopharyngeal and oropharyngeal samples for reverse transcription loop-mediated isothermal amplification (RT-LAMP). Additionally, the assay also utilizes the integration of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas12-based system using different guide RNAs of N, E and an internal control POP7 (human RNase P) genes along with visual detection via lateral flow readout-based dip sticks with unaided eye (~100 min). Overall, the clinical sensitivity and specificity of the CLEVER assay were 89.6% and 100%, respectively.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>Together, our CLEVER assay offers a point-of-care tool with no equipment dependency and minimum technical expertise requirement for COVID-19 diagnosis.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Significance and Impact of the Study<\/jats:title>\n                  <jats:p>To address the challenges associated with COVID-19 diagnosis, we need a faster, direct and more versatile detection method for an efficient epidemiological management of the COVID-19 outbreak. The present study involves developing a method for detection of SARS-CoV-2 in human body without RNA isolation step that can visually be detected with unaided eye. Taken together, our assay offers to overcome one major defect of the prior art, that is, RNA extraction step, which could limit the deployment of the previous assays in a testing site having limited lab infrastructure.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1111\/jam.15571","type":"journal-article","created":{"date-parts":[[2022,4,9]],"date-time":"2022-04-09T08:29:25Z","timestamp":1649492965000},"page":"410-421","source":"Crossref","is-referenced-by-count":30,"title":["CLEVER assay: A visual and rapid RNA extraction-free detection of SARS-CoV-2 based on CRISPR-Cas integrated RT-LAMP technology"],"prefix":"10.1093","volume":"133","author":[{"given":"Akansha","family":"Bhatt","sequence":"first","affiliation":[{"name":"Amity Institute of Biotechnology Amity University Haryana Gurugram Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeeshan","family":"Fatima","sequence":"additional","affiliation":[{"name":"Amity Institute of Biotechnology Amity University Haryana Gurugram Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Munindra","family":"Ruwali","sequence":"additional","affiliation":[{"name":"Amity Institute of Biotechnology Amity University Haryana Gurugram Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chitra Seetharam","family":"Misra","sequence":"additional","affiliation":[{"name":"CRISPR Biology Group, Applied Genomics Section Bhabha Atomic Research Centre Mumbai Maharashtra India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shyam Sunder","family":"Rangu","sequence":"additional","affiliation":[{"name":"CRISPR Biology Group, Applied Genomics Section Bhabha Atomic Research Centre Mumbai Maharashtra India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Devashish","family":"Rath","sequence":"additional","affiliation":[{"name":"CRISPR Biology Group, Applied Genomics Section Bhabha Atomic Research Centre Mumbai Maharashtra India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashok","family":"Rattan","sequence":"additional","affiliation":[{"name":"Pathkind Labs Gurugram Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6477-1870","authenticated-orcid":false,"given":"Saif","family":"Hameed","sequence":"additional","affiliation":[{"name":"Amity Institute of Biotechnology Amity University Haryana Gurugram Haryana India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,8,1]]},"reference":[{"key":"2023011711300280800_jam15571-bib-0001","doi-asserted-by":"crossref","DOI":"10.1016\/j.virusres.2020.198129","article-title":"iSCAN: An RT-LAMP-coupled CRISPR-Cas12 module for rapid, sensitive detection of SARS-CoV-2","volume":"288","author":"Ali","year":"2020","journal-title":"Virus Research"},{"key":"2023011711300280800_jam15571-bib-0002","article-title":"An inventory of diagnostic tools for detection of COVID-19","volume":"21","author":"Bhatt","year":"2021","journal-title":"Current Molecular Medicine"},{"issue":"7","key":"2023011711300280800_jam15571-bib-0003","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1038\/s41587-020-0513-4","article-title":"CRISPR-Cas12-based detection of SARS-CoV-2","volume":"38","author":"Broughton","year":"2020","journal-title":"Nature Biotechnology"},{"key":"2023011711300280800_jam15571-bib-0004","article-title":"Increased transmission of SARS-CoV-2 lineage B.1.1.7 (VOC 2020212\/01) is not accounted for by a replicative advantage in primary airway cells or antibody escape","author":"Brown","year":"2021","journal-title":"bioRxiv"},{"issue":"56","key":"2023011711300280800_jam15571-bib-0005","doi-asserted-by":"crossref","first-page":"6871","DOI":"10.1039\/D1CC02546E","article-title":"CRISPR\/Cas12a-mediated gold nanoparticle aggregation for colorimetric detection of SARS-CoV-2","volume":"57","author":"Cao","year":"2021","journal-title":"Chemical Communications (Cambridge, England)"},{"issue":"6387","key":"2023011711300280800_jam15571-bib-0006","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1126\/science.aar6245","article-title":"CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity","volume":"360","author":"Chen","year":"2018","journal-title":"Science (New York, N.Y.)"},{"issue":"3","key":"2023011711300280800_jam15571-bib-0007","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/s41579-018-0118-9","article-title":"Origin and evolution of pathogenic coronaviruses","volume":"17","author":"Cui","year":"2019","journal-title":"Nature Reviews. 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