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Rapid and simple detection methods are vital for early intervention to reduce severity and mortality. In this study, we designed highly specific RPA primers and crRNA (CRISPR RNA) based on the highly conserved CPSIT_0429 gene in the C. psittaci genome, and preliminarily established a nucleic acid detection method for C. psittaci using the RPA-CRISPR\/Cas12a system. In the two-step assay, the combination of the CPSIT_0429-F1\/R1 primer pair and CPSIT_0429-crRNA2 achieved a detection limit of 2\u00a0\u00d7\u00a010\u00b0 copies\/\u03bcL. Incorporating 20% glycerol enabled a one-tube assay with a limit of 2\u00a0\u00d7\u00a0102 copies\/\u03bcL. Furthermore, the method showed no cross-reactivity with common respiratory pathogens such as influenza virus, SARS-CoV-2, and Streptococcus pneumoniae, demonstrating excellent specificity. Both the two-step and one-tube methods were compared with qPCR-verified C. psittaci positive samples. The results indicated that both assays showed high consistency with qPCR results. The RPA-CRISPR\/Cas12a detection method is rapid, accurate, highly sensitive, and specific, providing a reliable platform for early diagnosis and clinical management of C. psittaci infections.<\/jats:p>","DOI":"10.1093\/lambio\/ovaf138","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T13:39:10Z","timestamp":1764941950000},"source":"Crossref","is-referenced-by-count":2,"title":["Nucleic acid detection method for\n                    <i>Chlamydia psittaci<\/i>\n                    based on RPA-CRISPR\/Cas12a"],"prefix":"10.1093","volume":"78","author":[{"given":"Qiong","family":"Li","sequence":"first","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Xu","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinyi","family":"Jiang","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Gong","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7634-4764","authenticated-orcid":false,"given":"Xujian","family":"Mao","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengming","family":"Wang","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Yao","sequence":"additional","affiliation":[{"name":"Pathogen Inspection Center, Changzhou Center for Disease Prevention and Control , 203 TaiShan Road, Changzhou 213022 Jiangsu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,12,8]]},"reference":[{"key":"2025122303423050300_bib1","doi-asserted-by":"publisher","first-page":"e0152123","DOI":"10.1128\/jcm.01521-23","article-title":"Retraction for Mustafa and Makhawi, \u201cSHERLOCK and DETECTR: cRISPR-Cas systems as potential rapid diagnostic tools for emerging infectious 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