{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T10:40:55Z","timestamp":1769856055828,"version":"3.49.0"},"reference-count":69,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,4,27]],"date-time":"2021-04-27T00:00:00Z","timestamp":1619481600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006109","name":"Vedeck\u00e1 Grantov\u00e1 Agent\u00fara M\u0160VVa\u0160 SR a SAV","doi-asserted-by":"publisher","award":["1\/0048\/16"],"award-info":[{"award-number":["1\/0048\/16"]}],"id":[{"id":"10.13039\/501100006109","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006109","name":"Vedeck\u00e1 Grantov\u00e1 Agent\u00fara M\u0160VVa\u0160 SR a SAV","doi-asserted-by":"publisher","award":["1\/0013\/20"],"award-info":[{"award-number":["1\/0013\/20"]}],"id":[{"id":"10.13039\/501100006109","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Chlamydia are Gram-negative, intracellular pathogens colonizing epithelial mucosa. They cause primarily atypical pneumonia and have recently been associated with chronic diseases. Diagnostics relies almost exclusively on serological methods; PCR tests are used rarely because in patients with positive ELISA, it is nearly impossible to identify chlamydial DNA. This paradox is associated with DNA degradation in sputum samples, low abundance, and low sensitivity of PCR systems. In a newly designed and validated \u201cnested\u201d PCR (NPCR) assay, it was possible to amplify DNA of Chlamydia known to infect humans in 31% samples. The reliability of the assay was confirmed by DNA sequencing, and all PCR products belonged exclusively to the Chlamydiales, mainly recognized as Chlamydia pneumoniae. Three samples were related to Ca. Rhabdochlamydia porcellionis and Ca. Renichlamydia lutjani, which infect arthropods. In one case, samples were taken from sick individual, indicating the potential as a human pathogen.<\/jats:p>","DOI":"10.3390\/microorganisms9050935","type":"journal-article","created":{"date-parts":[[2021,4,27]],"date-time":"2021-04-27T21:18:20Z","timestamp":1619558300000},"page":"935","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Reliable and Sensitive Nested PCR for the Detection of Chlamydia in Sputum"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5877-4904","authenticated-orcid":false,"given":"Martina","family":"Smolejov\u00e1","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Ilkovi\u010dova 6, 842 15 Bratislava, Slovakia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iveta","family":"Cihov\u00e1","sequence":"additional","affiliation":[{"name":"Department of Track and Field, Faculty of Physical Education and Sport, Comenius University, N\u00e1br. arm. gen. L. Svobodu 9, 814 69 Bratislava, Slovakia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1845-9031","authenticated-orcid":false,"given":"Pavol","family":"Sulo","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Ilkovi\u010dova 6, 842 15 Bratislava, Slovakia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.tim.2012.10.009","article-title":"Chlamydia pneumoniae: Modern insights into an ancient pathogen","volume":"21","author":"Roulis","year":"2013","journal-title":"Trends Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Persing, D., Tenover, F., Hayden, R., Ieven, M., Miller, M., Nolte, F., Tang, Y., and van Belkum, A. (2016). 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