{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T13:24:15Z","timestamp":1788269055543,"version":"build-2803163510"},"reference-count":70,"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"}],"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                      Urinary tract infections are one of the most common infectious diseases, with\n                      <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                      as the predominant pathogen. Traditional diagnostic methods fail to meet clinical demands for rapid and specific detection. Here, we developed an efficient urine\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      detection strategy via a reporter phage cocktail. Four reporter phages (T2::\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      , T4::\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      , T5::\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      , T6::\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      ) were constructed by the CRISPR\/Cas9 system combined with homologous recombination. One-step growth curves, optimal multiplicity of infection, and lytic efficiency showed that the\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      gene block insertion exerted heterogeneous effects on phages. Luminescence assays demonstrated that all five reporter phages (including previously preserved T7::\n                      <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                      ) and the cocktail offered favorable limits of detection (\u226510\n                      <jats:sup>3<\/jats:sup>\n                      CFU\/mL), high specificity, and no urine matrix interference. However, single phages exhibited limited coverage among 177 clinical\n                      <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                      isolates. But the reporter phage cocktail remedies this limitation. In large-scale clinical validation, the cocktail achieved sensitivity 73.15% (63.76%\u201381.22%), specificity 100.00% (99.53%\u2013100.00%), positive predictive value (PPV) 100.00% (95.44%\u2013100.00%), and negative predictive value (NPV) 96.42% (95.18%\u201397.36%) (all 95% confidence interval [CI]), and excellent concordance with the gold-standard method (Kappa = 0.83, 95% CI: 0.77\u20130.89), greatly outperforming single reporter phages (~40.00% sensitivity). This method requires no sample pretreatment, is simple to operate, and completes detection within 4 h, significantly improving diagnostic efficiency. Accordingly, it provides a novel platform for pathogen detection and supports the clinical translation of reporter phage diagnostics.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Urinary tract infections impose substantial economic and public health burdens. In this study, we successfully constructed\n                        <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                        -specific reporter phages T2::\n                        <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                        , T4::\n                        <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                        , T5::\n                        <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                        , and T6::\n                        <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                        . Combined with the previously preserved T7::\n                        <jats:italic toggle=\"yes\">Nluc<\/jats:italic>\n                        , these phages formed a reporter phage cocktail. Co-cultivation of this cocktail with clinical samples enabled rapid and specific detection of\n                        <jats:italic toggle=\"yes\">E. coli<\/jats:italic>\n                        in clinical urine, with a significantly shortened detection time (4 h) and good concordance with the gold-standard detection method (Kappa = 0.83), effectively improving detection efficiency and accuracy. This novel pathogen detection platform, integrating specific recognition and signal amplification, not only provides a new technical approach for the rapid and accurate diagnosis of clinical urinary tract infections but also effectively promotes the coordinated improvement of infectious disease diagnosis and treatment in terms of timeliness-precision-cost.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.00996-26","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T13:02:18Z","timestamp":1786021338000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Development and clinical validation of a CRISPR\/Cas9-engineered reporter phage cocktail for rapid detection of\n                    <i>Escherichia coli<\/i>\n                    in urine"],"prefix":"10.1128","volume":"14","author":[{"given":"Zhiyun","family":"Hao","sequence":"first","affiliation":[{"name":"PLA Medical School","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Zhao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Zhong","sequence":"additional","affiliation":[{"name":"PLA Medical School","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minwei","family":"Li","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6084-705X","authenticated-orcid":false,"given":"Chengbin","family":"Wang","sequence":"additional","affiliation":[{"name":"PLA Medical School","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2848-644X","authenticated-orcid":true,"given":"Chi","family":"Wang","sequence":"additional","affiliation":[{"name":"PLA Medical School","place":["Beijing, China"]},{"id":[{"id":"https:\/\/ror.org\/04gw3ra78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory Medicine, The First Medical Center of PLA General Hospital","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1080\/14787210.2025.2513459"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.3389\/fpubh.2022.888205"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-66865-8"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-57765-y"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/cmr.00129-24"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2025.117419"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2025.140415"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.euros.2022.08.009"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1039\/d1lc00888a"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-64608-3"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-39863-x"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2025.117907"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.watres.2025.124151"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.3724\/abbs.2025150"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2021.112973"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms13040745"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkad201"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkv188"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.03388-23"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/aac.00511-22"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1590\/0074-02760240204"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00125-13"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41522-024-00603-8"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhazmat.2025.140435"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2025.117830"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1021\/bp050201c"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSystems.00990-20"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.667332"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-75426-6"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.2c01822"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.3390\/v12090944"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.3390\/ph14070634"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2023.199272"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12866-023-02960-4"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2024.1347173"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2024.09.009"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2022.114378"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.70280"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae973"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12941-021-00470-7"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24087582"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12223-024-01205-9"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2011.11.015"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.3390\/ph14040347"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2022.05.016"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2015.07.014"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00442-20"},{"key":"e_1_3_3_49_2","doi-asserted-by":"publisher","DOI":"10.1093\/gbe\/evr059"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.ade9674"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.658374"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1119719109"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00573-18"},{"key":"e_1_3_3_54_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkz566"},{"key":"e_1_3_3_55_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02537"},{"key":"e_1_3_3_56_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1455710"},{"key":"e_1_3_3_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00211-21"},{"key":"e_1_3_3_58_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10111337"},{"key":"e_1_3_3_59_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12866-025-03895-8"},{"key":"e_1_3_3_60_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.09.015"},{"key":"e_1_3_3_61_2","doi-asserted-by":"publisher","DOI":"10.1039\/D4TB02178A"},{"key":"e_1_3_3_62_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.3c01569"},{"key":"e_1_3_3_63_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.2c03604"},{"key":"e_1_3_3_64_2","doi-asserted-by":"publisher","DOI":"10.3390\/s22103638"},{"key":"e_1_3_3_65_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2011.11.032"},{"key":"e_1_3_3_66_2","doi-asserted-by":"publisher","DOI":"10.3390\/v12060631"},{"key":"e_1_3_3_67_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00411-20"},{"key":"e_1_3_3_68_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2022.11.017"},{"key":"e_1_3_3_69_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-019-0437-z"},{"key":"e_1_3_3_70_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2025.1491961"},{"key":"e_1_3_3_71_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-68136-y"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00996-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00996-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T13:12:12Z","timestamp":1788268332000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.00996-26"}},"subtitle":[],"editor":[{"given":"Rosemary C.","family":"She","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,1]]},"references-count":70,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9,1]]}},"alternative-id":["10.1128\/spectrum.00996-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.00996-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,1]]},"assertion":[{"value":"2026-03-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-20","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-06","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00996-26"}}