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Although microscopy is the traditional diagnostic method for human filariasis, several polymerase chain reaction (PCR) methods have emerged as an alternative approach for identifying filarial parasites. The aim of this study is to compare three molecular methods and decide which is the most suitable for diagnosing human loiasis and mansonellosis in non\u2010endemic regions using dried blood spot (DBS) as a medium for sample collection and storage.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A total of 100 DBS samples, with their corresponding thin and thick blood smears, were selected for this study. Microscopy was used as the reference method to diagnose and calculate the microfilaraemia. Filarial DNA was extracted using the saponin\/Chelex method and the DNA isolated was assayed by Filaria\u2010real time\u2010PCR, filaria\u2010nested PCR, and cytochrome oxidase I PCR. All PCR products were subsequently purified and sequenced. The statistical values for each molecular test were calculated and compared.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Overall, 64 samples were identified as negative by all tests and a further 36 samples were positive by at least one of the methods used. The sensitivity and specificity were similar for the different molecular methods, all of which demonstrated good agreement with microscopy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Based on this study, and from a practical point of view (single and short amplification round), the optimal technique for diagnosing filarial infection in non\u2010endemic regions is filaria\u2010real time\u2010PCR, which presents high sensitivity and specificity and is also able to detect a wide range of human filariae.<\/jats:p><\/jats:sec>","DOI":"10.1111\/tmi.13786","type":"journal-article","created":{"date-parts":[[2022,6,2]],"date-time":"2022-06-02T18:07:12Z","timestamp":1654193232000},"page":"686-695","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Comparison of three <scp>PCR<\/scp>\u2010based methods to detect <i>Loa loa<\/i> and <i>Mansonella perstans<\/i> in long\u2010term frozen storage 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