{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T22:36:58Z","timestamp":1792795018179,"version":"4.5.0"},"reference-count":32,"publisher":"Microbiology Society","issue":"2","license":[{"start":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T00:00:00Z","timestamp":1770336000000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"research grant from the Children\u2019s Research Center of the University Children\u2019s Hospital Zurich","id":[{"id":"https:\/\/ror.org\/035vb3h42","id-type":"ROR","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100008464","name":"EMDO Stiftung","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100008464","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,2,6]]},"abstract":"<jats:p>\n                    <jats:bold>Introduction.<\/jats:bold>\n                    Direct detection of\n                    <jats:italic>Borrelia burgdorferi<\/jats:italic>\n                    by culture is considered the gold standard for confirming Lyme disease (LD). However,\n                    <jats:italic>B. burgdorferi<\/jats:italic>\n                    culture is not routinely used in clinical practice or research due to its lengthy protocol and low success rate. This study aimed to streamline the process by integrating a specific quantitative PCR (qPCR) screening early into the\n                    <jats:italic>B. burgdorferi<\/jats:italic>\n                    culture workflow for identification of cultures that are likely to yield viable spirochetes.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Methods.<\/jats:bold>\n                    Thirty-two blood plasma and 11 cerebrospinal fluid (CSF) samples were collected from 32 children with serologically confirmed LD and incubated in modified Kelly-Pettenkofer medium for up to 9\u2009weeks, with weekly assessments for viable spirochetes using microscopy. After 3\u2009weeks, the presence of\n                    <jats:italic>B. burgdorferi<\/jats:italic>\n                    DNA in culture was assessed by qPCR targeting the\n                    <jats:italic>B. burgdorferi flagellin B<\/jats:italic>\n                    gene. The estimated copy number of the target template was compared to the assay\u2019s 95% limit of detection (LOD).\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Results.<\/jats:bold>\n                    After 9\u2009weeks of incubation, viable spirochetes were observed in 2 (\n                    <jats:italic>n<\/jats:italic>\n                    =2\/32, 6.3%) plasma cultures and 3 (\n                    <jats:italic>n<\/jats:italic>\n                    =3\/11, 27.3%) CSF cultures. These were only observed in cultures showing copy numbers above 95% LOD in qPCR testing at week 3 (\n                    <jats:italic>n<\/jats:italic>\n                    =2\/3 plasma cultures, 66.7%;\n                    <jats:italic>n<\/jats:italic>\n                    =3\/3 CSF cultures, 100.0%).\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Conclusion.<\/jats:bold>\n                    Culturing\n                    <jats:italic>B. burgdorferi<\/jats:italic>\n                    is challenging and, despite a high workload, often not successful. qPCR may serve as an effective screening tool for\n                    <jats:italic>B. burgdorferi<\/jats:italic>\n                    cultures, enabling the culturing process to be streamlined by prioritizing cultures with target copy numbers exceeding the 95% LOD of the qPCR assay.\n                  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