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Molecular diagnostic techniques are not only faster and more sensitive than culture analysis, but can also detect a <jats:italic>Leptospira<\/jats:italic> infection before the appearance of antibodies. The aim of the present study was to analyze and compare two different PCR approaches applied to blood and urine specimens obtained from patients with clinical manifestations that were suggestive of leptospirosis. Furthermore, the results of these different PCR approaches were compared with the results of culture and serology analyses.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Results<\/jats:title>\n<jats:p>A total of 400 samples (234 blood or 58.5% and 166 urine of 41.5%) from 310 Slovenian patients with clinical manifestations suggestive of leptospirosis were tested using conventional PCR assays targeting the <jats:italic>rrs<\/jats:italic> gene and RT-PCR targeting the <jats:italic>lipL32<\/jats:italic> gene. Additionally, culture, serology and sequence analysis were performed for the majority of these samples. The PCR and RT-PCR results were concordant in 376 out of 400 of these samples (94.0%). Conventional PCR was positive for 27 out of 400 samples (6.8%) and RT-PCR was positive for 47 out of 400 samples (11.8%). Culture and microscopic agglutination tests supported these diagnoses.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Conclusions<\/jats:title>\n<jats:p>A comparison of the two PCR methods indicated that the RT-PCR targeting of the <jats:italic>lipL32<\/jats:italic> gene was faster, more sensitive and more specific for the determination of <jats:italic>Leptospira<\/jats:italic> DNA in these clinical samples.<\/jats:p>\n<\/jats:sec>","DOI":"10.1186\/s12866-020-01744-4","type":"journal-article","created":{"date-parts":[[2020,3,11]],"date-time":"2020-03-11T13:03:06Z","timestamp":1583931786000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":51,"title":["Evaluation of real-time PCR targeting the lipL32 gene for diagnosis of Leptospira infection"],"prefix":"10.1186","volume":"20","author":[{"given":"Da\u0161a","family":"Podgor\u0161ek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Ru\u017ei\u0107-Sablji\u0107","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mateja","family":"Logar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Pavlovi\u0107","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatjana","family":"Remec","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zvonko","family":"Baklan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emil","family":"Pal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4053-5590","authenticated-orcid":false,"given":"Tja\u0161a","family":"Cerar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,3,11]]},"reference":[{"issue":"2","key":"1744_CR1","doi-asserted-by":"publisher","first-page":"296","DOI":"10.1128\/CMR.14.2.296-326.2001","volume":"14","author":"PN Levett","year":"2001","unstructured":"Levett PN. 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Written informed consent was obtained from all patients. Adult subjects provided informed consent, and a parent or guardian of child participants provided informed consent on their behalf.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"We hereby declare that we have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"59"}}