{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T13:46:25Z","timestamp":1791812785939,"version":"4.3.4"},"reference-count":24,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T00:00:00Z","timestamp":1791763200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T00:00:00Z","timestamp":1791763200000},"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"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Lymphogranuloma venereum (LGV), caused by invasive\n                      <jats:italic toggle=\"yes\">Chlamydia trachomatis<\/jats:italic>\n                      genovars L1\u2013L4, requires timely differentiation from non-LGV strains to guide appropriate treatment and to strengthen case-based surveillance. We aimed to develop and validate a multiplex real-time PCR assay for simultaneous detection and differentiation of LGV and non-LGV\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      , targeting an LGV-associated chromosomal\n                      <jats:italic toggle=\"yes\">pmpH<\/jats:italic>\n                      deletion, an LGV-associated single-nucleotide polymorphism in the plasmid\n                      <jats:italic toggle=\"yes\">pgp3<\/jats:italic>\n                      gene, and the human\n                      <jats:italic toggle=\"yes\">RNaseP<\/jats:italic>\n                      gene as an internal control. Primers and probes were adapted from published assays and optimized for multiplex performance. Analytical validation was performed using synthetic gBlocks, cultured LGV and non-LGV strains, and Quality Control for Molecular Diagnostics (QCMD) proficiency panels. Clinical evaluation was conducted using 239\n                      <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                      NAAT-positive specimens submitted to the National Microbiology Laboratory for LGV detection and previously characterized using a\n                      <jats:italic toggle=\"yes\">pmpH-<\/jats:italic>\n                      based assay. All targets demonstrated efficient amplification (PCR efficiency ~100%) with no cross-reactivity. The limit of detection was 5\u201350 copies\/reaction, and analytical specificity for LGV\/non-LGV differentiation was 100%; results were fully concordant with QCMD panels. Clinical testing showed complete agreement with the original\n                      <jats:italic toggle=\"yes\">pmpH<\/jats:italic>\n                      assay for LGV-positive, non-LGV, and dual-positive specimens, while improving classification of indeterminate and previously\n                      <jats:italic toggle=\"yes\">pmpH<\/jats:italic>\n                      -negative samples. This assay enables improved, single-reaction LGV detection to support routine public health laboratory workflows and surveillance.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Accurate differentiation of lymphogranuloma venereum (LGV) from non-LGV\n                        <jats:italic toggle=\"yes\">Chlamydia trachomatis<\/jats:italic>\n                        is essential for appropriate treatment and effective surveillance. Current typing methods have reduced sensitivity in low-bacterial-load specimens, resulting in missed cases and reduced surveillance accuracy. We developed a multiplex real-time PCR assay that simultaneously detects and differentiates LGV and non-LGV\n                        <jats:italic toggle=\"yes\">C. trachomatis<\/jats:italic>\n                        using chromosomal and plasmid targets. This assay provides a streamlined and reliable approach for LGV detection. Implementation of the assay will improve turnaround time for actionable results, support appropriate treatment decisions, and reduce non-typeable results.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01461-26","type":"journal-article","created":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T13:00:59Z","timestamp":1791810059000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["A multiplex real-time PCR subtyping assay combining chromosomal (\n                    <i>pmpH<\/i>\n                    ) and plasmid (\n                    <i>pgp3<\/i>\n                    ) markers for differentiation of LGV and non-LGV\n                    <i>Chlamydia trachomatis<\/i>"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-6471-6491","authenticated-orcid":true,"given":"Sabaparvin","family":"Shaikh","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Beirnes","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harrison","family":"Deng","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]},{"id":[{"id":"https:\/\/ror.org\/02gfys938","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Microbiology and Infectious Diseases, Faculty of Health Sciences, University of Manitoba","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2568-6721","authenticated-orcid":true,"given":"Felicia","family":"Roy","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clarissa","family":"Klenke","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]},{"id":[{"id":"https:\/\/ror.org\/02gfys938","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Microbiology and Infectious Diseases, Faculty of Health Sciences, University of Manitoba","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kimberly","family":"Ta","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa","family":"Schulz","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]},{"id":[{"id":"https:\/\/ror.org\/02gfys938","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Microbiology and Infectious Diseases, Faculty of Health Sciences, University of Manitoba","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernard","family":"Abrenica","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quinton","family":"Murdock","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023xf2a37","id-type":"ROR","asserted-by":"publisher"}],"name":"National Microbiology Laboratory Branch, Public Health Agency of Canada","place":["Winnipeg, 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AIDS Programme of Research in South Africa (CAPRISA)","place":["Durban, South Africa"]},{"id":[{"id":"https:\/\/ror.org\/04qzfn040","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Microbiology, University of KwaZulu-Natal","place":["Durban, South Africa"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,10,12]]},"reference":[{"key":"e_1_3_4_2_2","unstructured":"Public Health Agency of Canada. 2023. Chlamydia Gonorrhea and Infectious Syphilis in Canada: 2021 Surveillance Data. Available from: https:\/\/www.canada.ca\/en\/public-health\/services\/publications\/diseases-conditions\/chlamydia-gonorrhea-infectious-syphilis-2021-surveillance-data.html. Retrieved 27 Jan 2026."},{"key":"e_1_3_4_3_2","unstructured":"Public Health Agency of Canada. 2021. Chlamydia and LGV guide: risk factors and clinical manifestations. Available from: https:\/\/www.canada.ca\/en\/public-health\/services\/infectious-diseases\/sexual-health-sexually-transmitted-infections\/canadian-guidelines\/chlamydia-lgv\/risk-factors-clinical-manifestation.html. 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