{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T17:26:19Z","timestamp":1785173179242,"version":"3.55.0"},"reference-count":27,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,5,8]],"date-time":"2025-05-08T00:00:00Z","timestamp":1746662400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2025,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Pulmonary lophomoniasis is an emerging disease caused by the protozoan pathogen <jats:italic>Lophomonas<\/jats:italic> spp. Recently, a conventional polymerase chain reaction (PCR) method has been developed. However, its sensitivity and specificity remain to be fully established. Therefore, this study aimed to develop in\u2010house conventional and multiplex PCR for the detection and identification of <jats:italic>Lophomonas<\/jats:italic> infections. Additionally, we attempted to compare the diagnostic performance of these novel PCR tests with the current microscopic examination method using BAL samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We studied 120 bronchoalveolar lavage (BAL) specimens of the patients clinically suspected of having lophomoniasis. The specimens were examined using three methods: microscopic examination (Giemsa staining), in\u2010house conventional PCR, and multiplex\u2010PCR. Moreover, multiplex\u2010PCR was used for the simultaneous identification of two species of <jats:italic>Lophomonas<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Out of the 120 BAL specimens tested, 30 (25%) tested positive through microscopic wet mount examination. Among the three techniques, multiplex\u2010PCR was the most sensitive (100%, 95% CI, 88.3\u2013100), while Giemsa staining had the lowest sensitivity (86.2%, 95% CI, 69.4\u201394.5). The data reveal a strong agreement between multiplex\u2010PCR and conventional PCR (\u03ba\u2009=\u20090.96), while the lowest agreement was found between multiplex\u2010PCR and microscopy methods (\u03ba\u2009=\u20090.16). The study also confirmed the presence of <jats:italic>L. blattarum<\/jats:italic> species in all samples using multiplex\u2010PCR.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This study demonstrates that the in\u2010house multiplex\u2010PCR is a robust and accurate diagnostic test for the detection and identification of <jats:italic>Lophomonas<\/jats:italic> species. Therefore, our findings suggest that this method may be a powerful tool to overcome some diagnostic pitfalls for lophomoniasis.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcla.70049","type":"journal-article","created":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T05:49:27Z","timestamp":1746769767000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development and Validation of In\u2010House Conventional and Multiplex <scp>PCR<\/scp> Methods for the Detection and Identification of <i>Lophomonas<\/i> spp.: An Innovative Approach"],"prefix":"10.1002","volume":"39","author":[{"given":"Maryam","family":"Nakhaei","sequence":"first","affiliation":[{"name":"Toxoplasmosis Research Center Communicable Diseases Institute, Mazandaran University of Medical Sciences  Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4690-6938","authenticated-orcid":false,"given":"Mahdi","family":"Fakhar","sequence":"additional","affiliation":[{"name":"Toxoplasmosis Research Center Communicable Diseases Institute, Mazandaran University of Medical Sciences  Sari Iran"},{"name":"Iranian National Registry Center for Lophomoniasis (INRCL) Imam Khomeini Hospital, Mazandaran University of Medical Sciences  Sari Iran"},{"name":"Department of Medical Microbiology and Immunology School of Medicine, Qom University of Medical Sciences  Qom Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abouzar","family":"Bagheri","sequence":"additional","affiliation":[{"name":"Molecular and Cell Biology Research Center, Faculty of Medicine Mazandaran University of Medical Sciences  Sari Iran"},{"name":"Immunogenetics Research Centre, Faculty of Medicine Mazandaran University of Medical Sciences  Sari 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University of Medical Sciences  Sari Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,5,8]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/jeu.12988"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12639-021-01381-2"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.2174\/1871526520666200727153142"},{"key":"e_1_2_12_5_1","volume-title":"Lophomonas and Lophomoniasis: Biology, Etiology, Epidemiology, Pathogenesis, Diagnosis and Treatment","author":"Fakhar 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