{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T11:09:00Z","timestamp":1782385740648,"version":"3.54.5"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2023,12,12]],"date-time":"2023-12-12T00:00:00Z","timestamp":1702339200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2023,12,12]],"date-time":"2023-12-12T00:00:00Z","timestamp":1702339200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["81974307"],"award-info":[{"award-number":["81974307"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"GDSTC | Natural Science Foundation of Guangdong Province","award":["2021A1515012255, 2023A1515012021"],"award-info":[{"award-number":["2021A1515012255, 2023A1515012021"]}]},{"name":"GDSTC | Natural Science Foundation of Guangdong Province","award":["2019A1515011827"],"award-info":[{"award-number":["2019A1515011827"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2023,12,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The occurrence of\n            <jats:italic>Chlamydia trachomatis<\/jats:italic>\n            (CT) and\n            <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n            (NG) is increasing worldwide, particularly in low- and middle-income countries. In cases where most infections are asymptomatic and remain in a coinfection condition. It may cause missed detections or the severe subsequent symptoms. Accurate diagnosis and convenient screening are essential for disease control. However, molecular diagnostics for CT\/NG predominantly rely on polymerase chain reaction (PCR)-based detection, which requires special devices and conditions. Here, we developed an isothermal CRISPR-based CT\/NG dual-target detection system by pairing recombinase polymerase amplification (RPA) and CRISPR-Cas12a\/13a detection. Multiplex RPA can amplify NG and CT simultaneously, and a single-pot reaction with the Cas12a\/13a can be performed for further recognizing the amplified target, driving the cleavage of the corresponding fluorescence reporter separately; thus, the signal can be monitored in different channels. This CRISPR-based CT\/NG detection system achieved analytical sensitivities of 10\u00b0 copies\/\u03bcL for both synthetic CT\/NG dsDNA and exhibited no cross-reaction with other species. In tests on 88 clinical samples, our CRISPR-based assay showed excellent agreement with the commercial assay Roche Cobas 4800 (100% accuracy for NG, 94.32% accuracy for CT, and 97.73% accuracy for CT\/NG coinfection) and showed higher positive percent agreement, negative percent agreement, and accuracy than those reported for TaqMan PCR in a previous study. In addition, the results of this CRISPR-based system could be acquired within 75 min. Our isothermal diagnostic method with promising performance provides more convenience than PCR-based methods for detection and screening.\n          <\/jats:p>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              A method for\n              <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n              (NG)\/\n              <jats:italic>Chlamydia trachomatis<\/jats:italic>\n              (CT) detection is developed using multiplex-recombinase polymerase amplification and Cas12a\/Cas13a. This method can detect NG and CT simultaneously with high sensitivity and specificity. This method has great potential to be further developed into larger-scale screening and point-of-care testing (POCT).\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.00464-23","type":"journal-article","created":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T13:01:58Z","timestamp":1698325318000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":11,"title":["An isothermal CRISPR-based diagnostic assay for\n            <i>Neisseria gonorrhoeae<\/i>\n            and\n            <i>Chlamydia trachomatis<\/i>\n            detection"],"prefix":"10.1128","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5463-9852","authenticated-orcid":true,"given":"Hao","family":"Luo","sequence":"first","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihong","family":"Zeng","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaona","family":"Yin","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuying","family":"Pan","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianjiang","family":"Yang","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingjing","family":"Liu","sequence":"additional","affiliation":[{"name":"Medical College, China Three Gorges University , Yichang, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolin","family":"Qin","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhanqin","family":"Feng","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4062-3865","authenticated-orcid":true,"given":"Wentao","family":"Chen","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9846-975X","authenticated-orcid":false,"given":"Heping","family":"Zheng","sequence":"additional","affiliation":[{"name":"Dermatology Hosptial, Southern Medical University , Guangzhou, China"},{"name":"Guangzhou Key Laboratory for Sexually Transmitted Diseases Control , Guangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.5534\/wjmh.190116"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0b013e318151497d"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.21101\/cejph.a5142"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0250871"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0000000000001446"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1111\/jdv.16913"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1136\/sextrans-2020-054515"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1136\/sextrans-2019-053999"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0000000000001403"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics7030059"},{"key":"e_1_3_3_13_2","unstructured":"Williams & Wilkins\n. 1989. Guide to clinical preventive services; an assessment of the effectiveness of 169 interventions"},{"key":"e_1_3_3_14_2","unstructured":"National Chlamydia Screening Programme (NCSP)\n. Available from: https:\/\/www.gov.uk\/government\/collections\/national-chlamydia-screening-programme-ncsp"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens9020091"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-51-12-1021"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1177\/0956462415618837"},{"key":"e_1_3_3_18_2","first-page":"1","article-title":"Recommendations for the laboratory-based detection of Chlamydia trachomatis and Neisseria gonorrhoeae","volume":"63","author":"Centers for disease control and prevention","year":"2014","unstructured":"Centers for disease control and prevention . 2014. Recommendations for the laboratory-based detection of Chlamydia trachomatis and Neisseria gonorrhoeae. 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