{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T14:26:57Z","timestamp":1788445617476,"version":"build-2803163510"},"reference-count":70,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,8]],"date-time":"2025-09-08T00:00:00Z","timestamp":1757289600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Agencia Nacional de Investigaci\u00f3n y Desarrollo (ANID)-Chile","award":["21251211"],"award-info":[{"award-number":["21251211"]}]},{"name":"FONCIENCIAS UDM010-Universidad del Magdalena","award":["21251211"],"award-info":[{"award-number":["21251211"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pneumonia caused by Pneumocystis jirovecii poses a serious threat, particularly to immunocompromised patients such as those with HIV\/AIDS, transplant recipients, or individuals undergoing chemotherapy. Its diagnosis is challenging because current methods, such as microscopy and certain molecular tests, have limitations in sensitivity and specificity, and require specialized equipment, which delays treatment initiation. In this context, CRISPR-Cas12-based methods offer a promising alternative: they are rapid, highly specific, sensitive, and low-cost, enabling more timely and accessible detection, even in resource-limited settings. We developed a simple and rapid detection platform based on the CRISPR-Cas12 coupled with lateral flow strips. A guide RNA was designed against DHPS, \u03b2-tubulin, and mtLSU rRNA genes. The guide corresponding to \u03b2-tubulin showed high sensitivity in the detection of P. jirovecii to produce a detectable fluorescence signal within the first 20\u201330 min. In addition, it demonstrated high specificity for P. jirovecii when DNA from other microorganisms was used. When coupled with lateral flow strips, high sensitivity and specificity were also observed for detecting positive samples, without the need for genetic amplification. CRISPR-Cas12 successfully detected P. jirovecii infection in an initial diagnostic application, demonstrating the potential of this method for integration into public health diagnostic systems, particularly in field, due to its adaptability, speed, and ease of use.<\/jats:p>","DOI":"10.3390\/ijms26178732","type":"journal-article","created":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T10:27:12Z","timestamp":1757500032000},"page":"8732","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["CRISPR-Cas12 Application for the Detection of Pneumocystis jirovecii in Immunodepression Patients Through Fluorescent and Lateral Flow Colorimetric Assay"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9172-1871","authenticated-orcid":false,"given":"Daniel","family":"Ulloa","sequence":"first","affiliation":[{"name":"Doctoral Program in Sciences, Mention in Applied Cellular and Molecular Biology, Universidad de La Frontera, Francisco Salazar Ave. 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Constanza","family":"N\u00fa\u00f1ez","sequence":"additional","affiliation":[{"name":"Biochemistry Program, Universidad de La Frontera, Francisco Salazar Ave. 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romina","family":"Matamala","sequence":"additional","affiliation":[{"name":"Biochemistry Program, Universidad de La Frontera, Francisco Salazar Ave. 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"An\u00edbal","family":"San Mart\u00edn","sequence":"additional","affiliation":[{"name":"Biotechnology Program, Universidad de La Frontera, Francisco Salazar Ave. 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5821-1518","authenticated-orcid":false,"given":"Dayana","family":"P\u00e1ez-De \u00c1vila","sequence":"additional","affiliation":[{"name":"Center for Genetics and Molecular Biology, Universidad del Magdalena, 29H3 Street No. 22-01, Santa Marta 470004, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7948-4145","authenticated-orcid":false,"given":"Jheyson","family":"Mercado-Vides","sequence":"additional","affiliation":[{"name":"Center for Genetics and Molecular Biology, Universidad del Magdalena, 29H3 Street No. 22-01, Santa Marta 470004, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3899-5993","authenticated-orcid":false,"given":"Juan","family":"Narv\u00e1ez","sequence":"additional","affiliation":[{"name":"Center for Genetics and Molecular Biology, Universidad del Magdalena, 29H3 Street No. 22-01, Santa Marta 470004, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Aguirre","sequence":"additional","affiliation":[{"name":"Center for Genetics and Molecular Biology, Universidad del Magdalena, 29H3 Street No. 22-01, Santa Marta 470004, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2339-2049","authenticated-orcid":false,"given":"Brian","family":"Effer","sequence":"additional","affiliation":[{"name":"Center of Excellence in Translational Medicine, Universidad de La Frontera, Francisco Salazar Ave. 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8426-9530","authenticated-orcid":false,"given":"Isabel","family":"Iturrieta-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Laboratory of Infectology and Clinical Immunology, Department of Preclinic Sciences, Medicine Faculty, Center of Excellence in Translational Medicine-Scientific and Technological Nucleus (CEMT-BIOREN), Universidad de La Frontera, Ave. Francisco Salazar 01145, Temuco 4811230, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Iturrieta-Gonz\u00e1lez, I., Chahin, C., Cabrera, J., Concha, C., Olivares-Ferretti, P., Briones, J., Vega, F., Bustos-Medina, L., and Fonseca-Salamanca, F. 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