{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T08:01:12Z","timestamp":1789200072108,"version":"build-2803163510"},"reference-count":26,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2022,3,21]],"date-time":"2022-03-21T00:00:00Z","timestamp":1647820800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Objectives<\/jats:title>\n<jats:p>Good-quality and sufficient DNA is essential for diagnostics and vaccine development. We aimed to compare six DNA extraction techniques applied to <jats:italic>Loa loa<\/jats:italic> microfilariae in order to evaluate the purity and integrity of extracts in terms of quality and quantity.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methods<\/jats:title>\n<jats:p>The microfilariae were purified via a Percoll gradient procedure with blood from hyper-microfilaremic individuals (&gt; 30,000 microfilaria [mf]\/ml). DNA extraction was carried out in duplicate at a rate of 350,000 mf\/tube for each technique: phenol\/chloroform, commercial Qiagen kit, salting out, Tris-EDTA, methanol, and cetyltrimethylammonium bromide (CTAB). The integrity, purity, concentration, and quality of the DNA extracts were successively verified by agarose gel electrophoresis, spectrophotometry (A<jats:sub>260<\/jats:sub>\/A<jats:sub>280<\/jats:sub> and A<jats:sub>260<\/jats:sub>\/A<jats:sub>230<\/jats:sub> wavelength ratio), Qubit fluorometry, and endonuclease and polymerase activity. The six techniques were compared on the basis of the following parameters: concentration, purity, efficiency, effectiveness, integrity, safety of the technique, as well as cost and duration of the protocol.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Results<\/jats:title>\n<jats:p>The ratios of the optical densities of the extracts A<jats:sub>260<\/jats:sub>\/A<jats:sub>280<\/jats:sub> and A<jats:sub>260<\/jats:sub>\/A<jats:sub>230<\/jats:sub> were, respectively: phenol\/chloroform (1.82; 1.11), Qiagen (1.93; 1.36), salting-out (1.9; 2.04), Tris-EDTA (1.99; 1.183), methanol (2.126; 1.343), and CTAB (2.01; 2.426). The DNA yield was: phenol\/chloroform (3.920 \u03bcg), Qiagen (10.280 \u03bcg), salting-out (10.390 \u03bcg), Tris-EDTA (0.5528 \u03bcg), methanol (0.1036 \u03bcg), and CTAB (1.115 \u03bcg). Endonuclease and polymerase activity was demonstrated by digestion of DNA and through amplicons obtained via polymerase chain reaction assays with phenol\/chloroform, Qiagen, and salting-out extracts.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>The phenol\/chloroform, Qiagen, and salting-out DNA extracts were all of good quality. Salting out had the best yield followed by Qiagen and then phenol\/chloroform. Endonuclease and polymerase activity was effective in all three extracts despite the presence of some contaminants. These methods are therefore suitable for the extraction of DNA from <jats:italic>Loa loa<\/jats:italic> microfilariae. Tris-EDTA and methanol did not show adequate sensitivity, while CTAB was found to be unsuitable.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0265582","type":"journal-article","created":{"date-parts":[[2022,3,21]],"date-time":"2022-03-21T17:25:34Z","timestamp":1647883534000},"page":"e0265582","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":11,"title":["Comparison of six methods for Loa loa genomic DNA extraction"],"prefix":"10.1371","volume":"17","author":[{"given":"Roland","family":"Dieki","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elsa-Rush","family":"Eyang-Assengone","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Makouloutou-Nzassi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F\u00e9licien","family":"Bangueboussa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edouard","family":"Nsi Emvo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8321-4500","authenticated-orcid":true,"given":"Jean Paul","family":"Akue","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2022,3,21]]},"reference":[{"issue":"9070","key":"pone.0265582.ref001","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/S0140-6736(96)11094-1","article-title":"Serious reactions after mass treatment of onchocerciasis with ivermectin in an area endemic for Loa loa infection","volume":"350","author":"J Gardon","year":"1997","journal-title":"Lancet"},{"issue":"3","key":"pone.0265582.ref002","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1016\/0035-9203(87)90163-5","article-title":"Filariasis due to Loa loa and Mansonella perstans: distribution in the region of Okondja, Haut-Ogoou\u00e9 Province, Gabon, with parasitological and serological follow-up over one year","volume":"81","author":"M Van Hoegaerden","year":"1987","journal-title":"Trans R Soc Trop Med Hyg"},{"issue":"5","key":"pone.0265582.ref003","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1016\/0035-9203(88)90218-0","article-title":"Common occurrence of amicrofilaraemic Loa loa filariasis within the endemic region","volume":"82","author":"A Dupont","year":"1988","journal-title":"Trans R Soc Trop Med Hyg"},{"issue":"5","key":"pone.0265582.ref004","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1111\/j.1365-3024.1992.tb00027.x","article-title":"Antibody-dependent cell-mediated immune reactions to Loa loa microfilariae in amicrofilaraemic subjects","volume":"14","author":"M Pinder","year":"1992","journal-title":"Parasite Immunol"},{"key":"pone.0265582.ref005","doi-asserted-by":"crossref","first-page":"109786","DOI":"10.1016\/j.mehy.2020.109786","article-title":"Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19","volume":"141","author":"J Kashir","year":"2020","journal-title":"Med Hypotheses"},{"issue":"4","key":"pone.0265582.ref006","doi-asserted-by":"crossref","first-page":"e94664","DOI":"10.1371\/journal.pone.0094664","article-title":"Development of a highly sensitive loop-mediated isothermal amplification (LAMP) method for the detection of Loa loa","volume":"9","author":"P. 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