{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T06:40:06Z","timestamp":1775198406111,"version":"3.50.1"},"reference-count":20,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,3,1]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n<jats:p>The present work evaluates sampling protocols, storage procedures, and DNA purification methods for <jats:italic>Leishmania<\/jats:italic> spp. detection and quantification in different biological samples. The efficiency of three preservation solutions, a phosphate buffer solution, an ethylenediaminetetraacetic acid (EDTA) buffer solution, and 70% ethanol, was compared in combination with three DNA extraction protocols: a commercial silica column kit, salting-out protein precipitation, and organic extraction with phenol-chloroform. Tissue samples from BALB\/c mice experimentally infected with <jats:italic>Leishmania <\/jats:italic>(<jats:italic>Leishmania<\/jats:italic>)<jats:italic> amazonensis<\/jats:italic>, <jats:italic>Leishmania <\/jats:italic>(<jats:italic>Viannia<\/jats:italic>)<jats:italic> braziliensis<\/jats:italic>, or <jats:italic>Leishmania <\/jats:italic>(<jats:italic>Leishmania<\/jats:italic>)<jats:italic> infantum<\/jats:italic> were stored in the three preservation solutions and subsequently subjected to the three different DNA extraction methods. The extracted DNA was then used in real-time polymerase chain reaction (PCR) assays for the detection and quantification of parasite ribosomal small subunit DNA targets as well as mammalian glyceraldehyde-3-phosphate dehydrogenase (<jats:italic>gapdh<\/jats:italic>) targets. The results of the optimized protocols showed that the DNA extraction method did not influence test quality, but DNA from samples preserved with the EDTA buffer solution produced higher amounts of target amplicons. Based on these results, we concluded that samples from suspected cases of leishmaniasis for submission to molecular diagnostic procedures should be preferentially preserved in EDTA, followed by any one of the DNA purification methods evaluated.<\/jats:p>","DOI":"10.4269\/ajtmh.21-1137","type":"journal-article","created":{"date-parts":[[2023,1,23]],"date-time":"2023-01-23T13:57:49Z","timestamp":1674482269000},"page":"548-554","source":"Crossref","is-referenced-by-count":1,"title":["Comparison of Sampling Procedures for the Molecular Diagnosis of Leishmaniases"],"prefix":"10.4269","volume":"108","author":[{"given":"Ricardo","family":"Andrade Zampieri","sequence":"first","affiliation":[{"name":"Department of Physiology, Institute of Biosciences, University of S\u00e3o Paulo, S\u00e3o Paulo, Brazil;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliana","family":"Ide Aoki","sequence":"additional","affiliation":[{"name":"Department of Physiology, Institute of Biosciences, University of S\u00e3o Paulo, S\u00e3o Paulo, Brazil;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl Erik","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Drammen Hospital, Vestre Viken Hospital Trust, Drammen, Norway;"},{"name":"Department of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, Norway;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey","family":"Jon Shaw","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, S\u00e3o Paulo, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucile","family":"Maria Floeter-Winter","sequence":"additional","affiliation":[{"name":"Department of Physiology, Institute of Biosciences, University of S\u00e3o Paulo, S\u00e3o Paulo, Brazil;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Lancet","first-page":"951","article-title":"Leishmaniasis","volume":"392","author":"Burza","year":"2018"},{"key":"b2","series-title":"Encyclopedia of Infection and Immunity","first-page":"622","author":"Antinori"},{"key":"b3","series-title":"Pathogens","first-page":"809","article-title":"One health approach to leishmaniases: understanding the disease dynamics through diagnostic tools","volume":"9","author":"Hong","year":"2020"},{"key":"b4","series-title":"An Bras Dermatol","first-page":"459","article-title":"Identification of Leishmania species by high-resolution DNA dissociation in cases of American cutaneous leishmaniasis","volume":"95","author":"Veasey","year":"2020"},{"key":"b5","series-title":"Clin Infect Dis","first-page":"e202","article-title":"Diagnosis and treatment of leishmaniasis: clinical practice guidelines by the Infectious Diseases Society of America 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