{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T12:08:17Z","timestamp":1784117297107,"version":"3.55.0"},"update-to":[{"DOI":"10.1371\/journal.pntd.0011715","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,10,31]],"date-time":"2023-10-31T00:00:00Z","timestamp":1698710400000}}],"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2023,10,19]],"date-time":"2023-10-19T00:00:00Z","timestamp":1697673600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Isfahan University of Medical Sciences, Isfahan, Iran","award":["3991123"],"award-info":[{"award-number":["3991123"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Background<\/jats:title>\n<jats:p>Developing more sensitive methods for the diagnosis of echinococcosis is essential. In this study PCR assay for sensitive detection of specific cell-free DNA (cfDNA) of <jats:italic>Echinococcus granulosus sensu lato<\/jats:italic> in the sera of the sheep naturally infected with echinococcosis was investigated.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methods<\/jats:title>\n<jats:p>To extract cfDNA from 35 infected sheep, the modified phenol-chloroform method was used for two different volumes (0.5 and 2 ml) of serum samples. From each extracted sample, two DNA volumes (5 and 10 \u03bcl) were amplified using both standard PCR and semi-nested PCR targeting NADH dehydrogenase subunit I.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Results<\/jats:title>\n<jats:p>Standard and semi-nested PCR on 0.5 ml of serum samples detected <jats:italic>Echinococcus<\/jats:italic> DNA in 8 and 12 out of 35 sheep, respectively; however, using 2 ml of serum samples, they detected 24 and 27 samples. By increasing the volume of template DNA, the PCRs could detect 29 and 33 out of 35 samples. The results were confirmed by sequencing of randomly selected PCR amplicons and comparing them with GenBank databases.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusions<\/jats:title>\n<jats:p>Larger volumes of serum for DNA extraction, greater volumes of DNA template for PCR, and employing a semi-nested PCR protocol, increased the sensitivity of PCR to 95%. This approach can also be applied to the diagnosis of echinococcosis in humans.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0011715","type":"journal-article","created":{"date-parts":[[2023,10,19]],"date-time":"2023-10-19T17:54:33Z","timestamp":1697738073000},"page":"e0011715","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Sensitive detection of specific cell-free DNA in serum samples from sheep with cystic echinococcosis"],"prefix":"10.1371","volume":"17","author":[{"given":"Mahboubeh","family":"Hadipour","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hossein 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