{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:11:45Z","timestamp":1787026305062,"version":"build-2736575974"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,6,15]],"date-time":"2022-06-15T00:00:00Z","timestamp":1655251200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100013262","name":"State Key Laboratory of Respiratory Disease","doi-asserted-by":"publisher","award":["SKLRD-Z-202224"],"award-info":[{"award-number":["SKLRD-Z-202224"]}],"id":[{"id":"10.13039\/100013262","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      can result in a serious pulmonary infection,\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      pneumonia, in immunocompetent hosts. The diagnosis of\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      pneumonia has long been a major clinical concern, and there are limitations with the currently utilized immunostaining and polymerase chain reaction diagnosis\/detection technologies (\n                      <jats:italic>e.g.<\/jats:italic>\n                      , insufficient sensitivity and accuracy). Hence, we sought to establish a rapid and RNA-specific transcription mediated amplification and CRISPR\/Cas13a-based diagnostics targeted\n                      <jats:italic>P. jirovecii<\/jats:italic>\n                      -mitochondrial large subunit ribosomal RNA.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>The procedure of the diagnostics included amplification of the extracted RNA samples by transcription mediated amplification, followed by CRISPR\/Cas13 detection, and ultimately, the judgment of the results after 30 minutes of fluorescence signal. Later, the diagnostic performance of the CRISPR\/Cas13-based diagnostics were tested on the 62 surplus clinical samples.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      This CRISPR\/Cas13-based diagnostics achieved limits of detection of approximately 2 copies\/\u00b5L transcribed RNA templates, with no cross reaction to other respiratory pathogens, including bacteria and fungi. Similar to in-house quantitative real-time polymerase chain reaction, CRISPR\/Cas13-based diagnostics was still positive in 243-fold diluted bronchial alveolar lavage fluid. A preliminary evaluation of 62 surplus bronchial alveolar lavage fluid samples from patients suspected of\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      pneumonia showed that CRISPR\/Cas13-based diagnostics achieved a 78.9% sensitivity and a 97.7% specificity in the diagnosis of\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      pneumonia.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Our study demonstrates that the CRISPR\/Cas13-based diagnostics technique has good performance for the accurate and specific diagnosis of\n                      <jats:italic>Pneumocystis jirovecii<\/jats:italic>\n                      pneumonia.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2022.904485","type":"journal-article","created":{"date-parts":[[2022,6,15]],"date-time":"2022-06-15T02:28:10Z","timestamp":1655260090000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":26,"title":["Development and Evaluation of Rapid and Accurate CRISPR\/Cas13-Based RNA Diagnostics for Pneumocystis jirovecii Pneumonia"],"prefix":"10.3389","volume":"12","author":[{"given":"Yangqing","family":"Zhan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoqing","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaoqiang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeqi","family":"Si","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanxiang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjun","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengtu","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Ye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,6,15]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.1111\/j.1469-0691.2010.03400.x","article-title":"Real-Time PCR Assay-Based Strategy for Differentiation Between Active Pneumocystis Jirovecii Pneumonia and Colonization in Immunocompromised Patients","volume":"17","author":"Alanio","year":"2011","journal-title":"Clin. 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