{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T01:36:47Z","timestamp":1788572207000,"version":"build-2803163510"},"reference-count":56,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2023,12,8]],"date-time":"2023-12-08T00:00:00Z","timestamp":1701993600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Indian Council of Medical Research, Ministry of Health and Family Welfare, Government of India"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Orientia tsutsugamushi is responsible for causing scrub typhus (ST) and is the leading cause of acute encephalitis syndrome (AES) in AES patients. A rapid and sensitive method to detect scrub typhus on-site is essential for the timely deployment of control measures. In the current study, we developed a rapid, sensitive, and instrument-free lateral flow assay (LFA) detection method based on CRISPR\/Cas12a technology for diagnosing ST (named LoCIST). The method is completed in three steps: first, harnessing the ability of recombinase polymerase for isothermal amplification of the target gene; second, CRISPR\/Cas12a-based recognition of the target; and third, end-point detection by LFA. The detection limit of LoCIST was found to be one gene copy of ST genomic DNA per reaction, and the process was complete within an hour. In 81 clinical samples, the assay showed no cross-reactivity with other rickettsial DNA and was 100% consistent with PCR detection of ST. LoCIST demonstrated 97.6% sensitivity and 100% specificity. Overall, the LoCIST offers a novel alternative for the portable, simple, sensitive, and specific detection of ST, and it may help prevent and control AES outbreaks due to ST. In conclusion, LoCIST does not require specialized equipment and poses a potential for future applications as a point-of-care diagnostic.<\/jats:p>","DOI":"10.3390\/bios13121021","type":"journal-article","created":{"date-parts":[[2023,12,11]],"date-time":"2023-12-11T03:25:31Z","timestamp":1702265131000},"page":"1021","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["CRISPR\/Cas12a-Based Detection Platform for Early and Rapid Diagnosis of Scrub Typhus"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7497-0291","authenticated-orcid":false,"given":"Pooja","family":"Bhardwaj","sequence":"first","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8843-4297","authenticated-orcid":false,"given":"Nikita Shrikant","family":"Nanaware","sequence":"additional","affiliation":[{"name":"ICMR-National AIDS Research Institute, Bhosari, Pune 411026, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1517-5757","authenticated-orcid":false,"given":"Sthita Pragnya","family":"Behera","sequence":"additional","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Smita","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"ICMR-National AIDS Research Institute, Bhosari, Pune 411026, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hirawati","family":"Deval","sequence":"additional","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3965-395X","authenticated-orcid":false,"given":"Rajesh","family":"Kumar","sequence":"additional","affiliation":[{"name":"RGSC, Department of Genetics and Plant Breeding, Banaras Hindu University, Varanasi 221005, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4704-7116","authenticated-orcid":false,"given":"Gaurav Raj","family":"Dwivedi","sequence":"additional","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajni","family":"Kant","sequence":"additional","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7597-5601","authenticated-orcid":false,"given":"Rajeev","family":"Singh","sequence":"additional","affiliation":[{"name":"ICMR-Regional Medical Research Centre Gorakhpur, BRD Medical College Campus, Gorakhpur 273013, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Xu, G., Walker, D.H., Jupiter, D., Melby, P.C., and Arcari, C.M. 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