{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T13:23:36Z","timestamp":1788269016037,"version":"build-2803163510"},"reference-count":45,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"the Key Research and Development Program of Yunnan Province","award":["202403AC100011"],"award-info":[{"award-number":["202403AC100011"]}]},{"name":"First-Class Discipline Team of Kunming Medical University","award":["2024XKTDYS01"],"award-info":[{"award-number":["2024XKTDYS01"]}]},{"name":"Project of AIDS Bureau of Yunnan Province"},{"name":"Xingdian Talents Support Project of Yunnan Province","award":["RLXZ20230001"],"award-info":[{"award-number":["RLXZ20230001"]}]},{"name":"Yunnan Province Clinical Center for Skin Immune Diseases","award":["YWLCYXZX2023300076"],"award-info":[{"award-number":["YWLCYXZX2023300076"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Chlamydia trachomatis<\/jats:italic>\n                      (\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      ) and\n                      <jats:italic toggle=\"yes\">Neisseria gonorrhoeae<\/jats:italic>\n                      (\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      ) are common sexually transmitted pathogens. Although frequently asymptomatic, they can cause severe reproductive complications (e.g., epididymitis, salpingitis, pelvic inflammatory disease, and infertility) if untreated. We developed a duplex high-fidelity loop-mediated isothermal amplification (HiFi-LAMP) assay for simultaneous detection of\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      . The assay demonstrated high sensitivity and specificity, with limits of detection (LODs) of 358 and 30 copies per 25 \u03bcL reaction for\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      , respectively. The performance of the assays was evaluated by comparing with the commercial singleplex qPCR assays using 156 paired urine and genital secretion samples. The assays have 100% specificity across all samples and achieved 100% sensitivity for the detection of\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      in urine and\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      in genital secretion samples and 98.7% and 88.5% sensitivity for\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      in genital secretions and\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      in urines, respectively. Extraction-free HiFi-LAMP assays directly using a small amount of simply treated urine were established and showed 87.9% sensitivity and 100% specificity for the detection of\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      in 70 urine samples. The new HiFi-LAMP assays can detect all\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      - and\/or\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      -positive samples within 30 min, implying that the entire HiFi-LAMP assay (including sample process) can be completed within 40 min, significantly shorter than about 90 min by the qPCR assays. The new HiFi-LAMP assays provide a rapid, simple, and cost-effective point-of-care testing (POCT) tool for clinical detection and\/or surveillance of\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                      in resource-limited settings.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Asymptomatic\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        infections significantly increase the risk of silent transmission.\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        were cocirculating in sexually active individuals. Of 156 individuals, we detected 7 (4.5%) individuals positive for both\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        , implying an urgent need for rapid accurate and duplex POCT assay for timely detection of\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        . The development of a duplex HiFi-LAMP assay enables the simultaneous detection of\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        with high sensitivity and specificity. The assay offers high diagnostic concordance with commercial qPCR assays and has lower cost and shorter reaction time. Given the availability of various portable fluorescent thermal cyclers and\/or isothermal fluorimeters, the development of extraction-free HiFi-LAMP assay directly using simply treated urine will lower the barriers of sexually active individuals to routine screening and facilitate clinical detection and surveillance of\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        and\n                        <jats:italic toggle=\"yes\">N. gonorrhoeae<\/jats:italic>\n                        in resource-limited settings.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03416-25","type":"journal-article","created":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T13:00:17Z","timestamp":1784898017000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Duplex, extraction-free, HiFi-LAMP assay for point-of-care detection of\n                    <i>Chlamydia trachomatis<\/i>\n                    and\n                    <i>Neisseria gonorrhoeae<\/i>"],"prefix":"10.1128","volume":"14","author":[{"given":"Zhengfang","family":"Wang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/038c3w259","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Dermatology and Venereology, First A\ufb03liated Hospital of Kunming Medical University","place":["Kunming, People's Republic of China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongjuan","family":"Zhao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/013q1eq08","id-type":"ROR","asserted-by":"publisher"}],"name":"Shanghai Public Health Clinical Center, Fudan University","place":["Shanghai, People's Republic of 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