{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T12:34:31Z","timestamp":1792154071588,"version":"4.4.0"},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T00:00:00Z","timestamp":1643328000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["C.B is senior investigator"],"award-info":[{"award-number":["C.B is senior investigator"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["Scholarship"],"award-info":[{"award-number":["Scholarship"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["resjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Medical Vet Entomology"],"published-print":{"date-parts":[[2022,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The study aimed to develop a multiplex qPCR to detect\n                    <jats:italic>Leishmania infantum<\/jats:italic>\n                    load in different sandfly sample settings using\n                    <jats:italic>Leishmania<\/jats:italic>\n                    kDNA and sandfly vacuolar ATPase (VATP) subunit C as internal control gene. The amplification of\n                    <jats:italic>Lutzomyia longipalpis<\/jats:italic>\n                    VATP gene was evaluated together with\n                    <jats:italic>Leishmania infantum<\/jats:italic>\n                    kDNA in a multiplex reaction. The concentration of VATP gene oligonucleotides was adjusted until no statistically significant difference was observed between all multiplex standard curves and singleplex curves, that is, only kDNA amplification. Limit of detection (LoD) was measured using a probit model and a cut\u2010off defined by receiver operating characteristic analysis. Limit of quantification (LoQ) was assessed by a linear model using the coefficient of variation threshold of 25%. After assuring VATP gene amplification, its primer\u2013probe concentrations were best at 100\u2009nM\/10\u00a0nM, respectively. The cut\u2010off\n                    <jats:italic>C<\/jats:italic>\n                    <jats:sub>q<\/jats:sub>\n                    value for\n                    <jats:italic>L. infantum<\/jats:italic>\n                    kDNA was defined as 35.46 with 100% of sensitivity and specificity. A total of 95% LoD was determined to be of 0.162 parasites while LoQ was 5.858. Our VATP\/kDNA multiplex qPCR assay shows that it can be used to evaluate both DNA integrity and determine\n                    <jats:italic>L. infantum<\/jats:italic>\n                    load in\n                    <jats:italic>L. longipalpis<\/jats:italic>\n                    even for low yielded samples, that is, individual midguts.\n                  <\/jats:p>","DOI":"10.1111\/mve.12564","type":"journal-article","created":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T10:14:06Z","timestamp":1643364846000},"page":"176-184","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Multiplex\n                    <scp>qPCR<\/scp>\n                    assay to determine\n                    <i>Leishmania infantum<\/i>\n                    load in\n                    <i>Lutzomyia longipalpis<\/i>\n                    sandfly samples"],"prefix":"10.1111","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6575-8208","authenticated-orcid":false,"given":"Tiago Feitosa","family":"Mota","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o Parasito\u2010Hospedeiro e Epidemiologia (LaIPHE), Instituto Gon\u00e7alo Moniz (IGM) FIOCRUZ\u2010BA, Rua Waldemar Falc\u00e3o  Salvador Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3367-5365","authenticated-orcid":false,"given":"Claudia Ida","family":"Brodskyn","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o Parasito\u2010Hospedeiro e Epidemiologia (LaIPHE), Instituto Gon\u00e7alo Moniz (IGM) FIOCRUZ\u2010BA, Rua Waldemar Falc\u00e3o  Salvador Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6438-8284","authenticated-orcid":false,"given":"Luis 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