{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T10:05:49Z","timestamp":1790762749558,"version":"4.1.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"4","license":[{"start":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T00:00:00Z","timestamp":1641772800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T00:00:00Z","timestamp":1641772800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPPGH5342"],"award-info":[{"award-number":["OPPGH5342"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]},{"name":"BONFOR"},{"name":"Deutsches Zentrum f\u00fcr Infektionsforschung, Clinical Leave Stipend","award":["TI 07.001_Schlabe_00"],"award-info":[{"award-number":["TI 07.001_Schlabe_00"]}]},{"name":"Deutsches Zentrum f\u00fcr Infektionsforschung, Clinical Leave Stipend","award":["TI 07.001_Schlabe_01"],"award-info":[{"award-number":["TI 07.001_Schlabe_01"]}]},{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["TI 03.907"],"award-info":[{"award-number":["TI 03.907"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["TI 03.907"],"award-info":[{"award-number":["TI 03.907"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["TI 03.907"],"award-info":[{"award-number":["TI 03.907"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["EXC2151 \u2013 390873048"],"award-info":[{"award-number":["EXC2151 \u2013 390873048"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Universit\u00e4tsklinikum Bonn"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasitol Res"],"published-print":{"date-parts":[[2022,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    The filarial nematode\n                    <jats:italic>Onchocerca volvulus<\/jats:italic>\n                    causes onchocerciasis (river blindness), a neglected tropical disease affecting 21 million people, mostly in Sub-Saharan Africa. Targeting the endosymbiont\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    with antibiotics leads to permanent sterilization and killing of adult worms. The gold standard to assess\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    depletion is the histological examination of adult worms in nodules beginning at 6\u00a0months post-treatment. However, nodules can only be used once, limiting the time points to monitor\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    depletion. A diagnostic to longitudinally monitor\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    depletion from microfilariae (MF) at more frequent intervals\u2009&lt;\u20096\u00a0months post-treatment would accelerate clinical trials of antiwolbachials. We developed a TaqMan qPCR amplifying the single-copy gene\n                    <jats:italic>w<\/jats:italic>\n                    Ov\n                    <jats:italic>ftsZ<\/jats:italic>\n                    to quantify\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    from as few as one MF that had migrated from skin biopsies and compared quantification using circular and linearized plasmids or synthetic dsDNA (gBlock\u00ae). qPCR for MF from the rodent nematode\n                    <jats:italic>Litomosoides sigmodontis<\/jats:italic>\n                    was used to support the reproducibility and validate the principle. The qPCR using as few as 2 MF from\n                    <jats:italic>O. volvulus<\/jats:italic>\n                    and\n                    <jats:italic>L. sigmodontis<\/jats:italic>\n                    reproducibly quantified\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    . Use of a linearized plasmid standard or synthesized dsDNA resulted in numbers of\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    \/MF congruent with biologically plausible estimates in\n                    <jats:italic>O. volvulus<\/jats:italic>\n                    and\n                    <jats:italic>L. sigmodontis<\/jats:italic>\n                    MF. The qPCR assay yielded a median of 48.8 (range 1.5\u2013280.5)\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    \/\n                    <jats:italic>O. volvulus<\/jats:italic>\n                    MF. The qPCR is a sensitive tool for quantifying\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    in a few MF from skin biopsies and allows for establishing the qPCR as a surrogate parameter for monitoring\n                    <jats:italic>Wolbachia<\/jats:italic>\n                    depletion in adult worms of new antiwolbachial candidates.\n                  <\/jats:p>","DOI":"10.1007\/s00436-021-07411-5","type":"journal-article","created":{"date-parts":[[2022,1,10]],"date-time":"2022-01-10T08:04:45Z","timestamp":1641801885000},"page":"1199-1206","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A qPCR to quantify Wolbachia from few Onchocerca volvulus microfilariae as a surrogate for adult worm histology in clinical trials of antiwolbachial drugs"],"prefix":"10.1007","volume":"121","author":[{"given":"Stefan","family":"Schlabe","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Korir","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"L\u00e4mmer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederic","family":"Landmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bettina","family":"Dubben","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marianne","family":"Koschel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Albers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda Batsa","family":"Debrah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander Yaw","family":"Debrah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc P.","family":"H\u00fcbner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3096-2465","authenticated-orcid":false,"given":"Kenneth","family":"Pfarr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ute","family":"Klarmann-Schulz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Achim","family":"Hoerauf","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,1,10]]},"reference":[{"key":"7411_CR1","unstructured":"Albers AC (2011) Evaluation of Wolbachia depletion after antibiotic treatment using real-time PCR in onchocerciasis and lymphatic filariasis, and analysis of genetic associations in lymphatic filariasis. Rheinische Friedrich Wilhelms Universit\u00e4t Bonn"},{"key":"7411_CR2","unstructured":"Batsa-Debrah L et al (2017) Doxycycline for the treatment of onchocerciasis: a daily dose of 100 mg for 6 weeks shows reduction of fertile female Onchocera volvulus equivalent to 200 mg\/d (531). Paper presented at the 66th Annual Meeting ASTMH, Baltimore, Maryland, USA, November 5\u20139, 2017"},{"issue":"4","key":"7411_CR3","doi-asserted-by":"publisher","first-page":"933","DOI":"10.1093\/cid\/ciz889","volume":"71","author":"L Batsa Debrah","year":"2020","unstructured":"Batsa Debrah L et al (2020) Comparison of repeated doses of ivermectin versus ivermectin plus albendazole for the treatment of onchocerciasis: a randomized, open-label, clinical trial. Clin Infect Dis 71(4):933\u2013943. https:\/\/doi.org\/10.1093\/cid\/ciz889","journal-title":"Clin Infect Dis"},{"issue":"1","key":"7411_CR4","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1186\/1475-2883-2-10","volume":"2","author":"DW B\u00fcttner","year":"2003","unstructured":"B\u00fcttner DW, Wanji S, Bazzocchi C, Bain O, Fischer P (2003) Obligatory symbiotic Wolbachia endobacteria are absent from Loa loa. Filaria J 2(1):10. https:\/\/doi.org\/10.1186\/1475-2883-2-10","journal-title":"Filaria J"},{"issue":"4","key":"7411_CR5","doi-asserted-by":"publisher","first-page":"177","DOI":"10.2144\/btn-2018-2000","volume":"64","author":"J Conte","year":"2018","unstructured":"Conte J, Potoczniak MJ, Tobe SS (2018) Using synthetic oligonucleotides as standards in probe-based qPCR. Biotechniques 64(4):177\u2013179. https:\/\/doi.org\/10.2144\/btn-2018-2000","journal-title":"Biotechniques"},{"issue":"4","key":"7411_CR6","doi-asserted-by":"publisher","first-page":"517","DOI":"10.1093\/cid\/civ363","volume":"61","author":"AY Debrah","year":"2015","unstructured":"Debrah AY et al (2015) Doxycycline leads to sterility and enhanced killing of female Onchocerca volvulus worms in an area with persistent microfilaridermia after repeated ivermectin treatment: a randomized, placebo-controlled, double-blind trial. Clin Infect Dis 61(4):517\u2013526. https:\/\/doi.org\/10.1093\/cid\/civ363","journal-title":"Clin Infect Dis"},{"issue":"3","key":"7411_CR7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007218","volume":"13","author":"N Fattouh","year":"2019","unstructured":"Fattouh N, Cazevieille C, Landmann F (2019) Wolbachia endosymbionts subvert the endoplasmic reticulum to acquire host membranes without triggering ER stress. PLoS Negl Trop Dis 13(3):e0007218. https:\/\/doi.org\/10.1371\/journal.pntd.0007218","journal-title":"PLoS Negl Trop Dis"},{"issue":"9266","key":"7411_CR8","doi-asserted-by":"publisher","first-page":"1415","DOI":"10.1016\/S0140-6736(00)04581-5","volume":"357","author":"A Hoerauf","year":"2001","unstructured":"Hoerauf A, Mand S, Adjei O, Fleischer B, Buttner DW (2001) Depletion of Wolbachia endobacteria in Onchocerca volvulus by doxycycline and microfilaridermia after ivermectin treatment. Lancet 357(9266):1415\u20131416. https:\/\/doi.org\/10.1016\/S0140-6736(00)04581-5","journal-title":"Lancet"},{"issue":"3","key":"7411_CR9","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1007\/s00430-007-0062-1","volume":"197","author":"A Hoerauf","year":"2008","unstructured":"Hoerauf A et al (2008) Wolbachia endobacteria depletion by doxycycline as antifilarial therapy has macrofilaricidal activity in onchocerciasis: a randomized placebo-controlled study. Med Microbiol Immunol 197(3):295\u2013311. https:\/\/doi.org\/10.1007\/s00430-007-0062-1","journal-title":"Med Microbiol Immunol"},{"issue":"4","key":"7411_CR10","doi-asserted-by":"publisher","first-page":"1414","DOI":"10.1073\/pnas.1816585116","volume":"116","author":"WD Hong","year":"2019","unstructured":"Hong WD et al (2019) AWZ1066S, a highly specific anti-Wolbachia drug candidate for a short-course treatment of filariasis. Proc Natl Acad Sci U S A 116(4):1414\u20131419. https:\/\/doi.org\/10.1073\/pnas.1816585116","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"3","key":"7411_CR11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0009545","volume":"5","author":"Y Hou","year":"2010","unstructured":"Hou Y, Zhang H, Miranda L, Lin S (2010) Serious overestimation in quantitative PCR by circular (supercoiled) plasmid standard: microalgal PCNA as the model gene. PLoS ONE 5(3):e9545. https:\/\/doi.org\/10.1371\/journal.pone.0009545","journal-title":"PLoS ONE"},{"issue":"8","key":"7411_CR12","doi-asserted-by":"publisher","first-page":"e0007636","DOI":"10.1371\/journal.pntd.0007636","volume":"13","author":"MP H\u00fcbner","year":"2019","unstructured":"H\u00fcbner MP et al (2019) In vivo kinetics of Wolbachia depletion by ABBV-4083 in L. sigmodontis adult worms and microfilariae. PLoS Negl Trop Dis 13(8):e0007636. https:\/\/doi.org\/10.1371\/journal.pntd.0007636","journal-title":"PLoS Negl Trop Dis"},{"issue":"7","key":"7411_CR13","doi-asserted-by":"publisher","first-page":"e0008427","DOI":"10.1371\/journal.pntd.0008427","volume":"14","author":"MP H\u00fcbner","year":"2020","unstructured":"H\u00fcbner MP et al (2020) Oxfendazole mediates macrofilaricidal efficacy against the filarial nematode Litomosoides sigmodontis in vivo and inhibits Onchocerca spec. motility in vitro. PLoS Negl Trop Dis 14(7):e0008427. https:\/\/doi.org\/10.1371\/journal.pntd.0008427","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"7411_CR14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0005156","volume":"11","author":"U Klarmann-Schulz","year":"2017","unstructured":"Klarmann-Schulz U et al (2017) Comparison of doxycycline, minocycline, doxycycline plus albendazole and albendazole alone in their efficacy against onchocerciasis in a randomized, open-label, pilot trial. PLoS Negl Trop Dis 11(1):e0005156. https:\/\/doi.org\/10.1371\/journal.pntd.0005156","journal-title":"PLoS Negl Trop Dis"},{"key":"7411_CR15","doi-asserted-by":"publisher","unstructured":"Kr\u00fccken J et al (2021) Development of emodepside as possible adulticidal treatment for human onchocerciasis \u2013 the fruit of a successful industrial-academic collaboration. PLoS Pathog 17(7):e1009682. https:\/\/doi.org\/10.1371\/journal.ppat.1009682","DOI":"10.1371\/journal.ppat.1009682"},{"key":"7411_CR16","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1186\/1756-3305-5-16","volume":"5","author":"F Landmann","year":"2012","unstructured":"Landmann F, Foster JM, Slatko BE, Sullivan W (2012) Efficient in vitro RNA interference and immunofluorescence-based phenotype analysis in a human parasitic nematode Brugia Malayi. Parasit Vectors 5:16. https:\/\/doi.org\/10.1186\/1756-3305-5-16","journal-title":"Parasit Vectors"},{"issue":"12","key":"7411_CR17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0029101","volume":"6","author":"CH Lin","year":"2011","unstructured":"Lin CH, Chen YC, Pan TM (2011) Quantification bias caused by plasmid DNA conformation in quantitative real-time PCR assay. PLoS ONE 6(12):e29101. https:\/\/doi.org\/10.1371\/journal.pone.0029101","journal-title":"PLoS ONE"},{"issue":"6","key":"7411_CR18","doi-asserted-by":"publisher","first-page":"1007","DOI":"10.2307\/3279835","volume":"63","author":"A Martinez-Palomo","year":"1977","unstructured":"Martinez-Palomo A, Martinez-Baez M (1977) Ultrastructure of the microfilaria of Onchocerca volvulus from Mexico. J Parasitol 63(6):1007\u20131018. https:\/\/doi.org\/10.2307\/3279835","journal-title":"J Parasitol"},{"issue":"9","key":"7411_CR19","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002922","volume":"8","author":"LR Serbus","year":"2012","unstructured":"Serbus LR et al (2012) A cell-based screen reveals that the albendazole metabolite, albendazole sulfone, targets Wolbachia. PLoS Pathog 8(9):e1002922. https:\/\/doi.org\/10.1371\/journal.ppat.1002922","journal-title":"PLoS Pathog"},{"issue":"1","key":"7411_CR20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0006116","volume":"12","author":"S Specht","year":"2018","unstructured":"Specht S et al (2018) Combinations of registered drugs reduce treatment times required to deplete Wolbachia in the Litomosoides sigmodontis mouse model. PLoS Negl Trop Dis 12(1):e0006116. https:\/\/doi.org\/10.1371\/journal.pntd.0006116","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"7411_CR21","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1007\/s00436-009-1624-5","volume":"106","author":"S Specht","year":"2009","unstructured":"Specht S, Hoerauf A, Adjei O, Debrah A, Buttner DW (2009) Newly acquired Onchocerca volvulus filariae after doxycycline treatment. Parasitol Res 106(1):23\u201331. https:\/\/doi.org\/10.1007\/s00436-009-1624-5","journal-title":"Parasitol Res"},{"key":"7411_CR22","doi-asserted-by":"publisher","unstructured":"Taylor MJ, et al. (2019) Preclinical development of an oral anti-Wolbachia macrolide drug for the treatment of lymphatic filariasis and onchocerciasis. Sci Transl Med 11(483), https:\/\/doi.org\/10.1126\/scitranslmed.aau2086","DOI":"10.1126\/scitranslmed.aau2086"},{"issue":"4","key":"7411_CR23","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0002169","volume":"7","author":"HC Turner","year":"2013","unstructured":"Turner HC, Churcher TS, Walker M, Osei-Atweneboana MY, Prichard RK, Basanez MG (2013) Uncertainty surrounding projections of the long-term impact of ivermectin treatment on human onchocerciasis. PLoS Negl Trop Dis 7(4):e2169. https:\/\/doi.org\/10.1371\/journal.pntd.0002169","journal-title":"PLoS Negl Trop Dis"},{"issue":"8","key":"7411_CR24","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.1086\/501351","volume":"42","author":"JD Turner","year":"2006","unstructured":"Turner JD et al (2006) A randomized, double-blind clinical trial of a 3-week course of doxycycline plus albendazole and ivermectin for the treatment of Wuchereriabancrofti infection. Clin Infect Dis 42(8):1081\u20131089. https:\/\/doi.org\/10.1086\/501351","journal-title":"Clin Infect Dis"},{"issue":"2","key":"7411_CR25","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007159","volume":"13","author":"TW von Geldern","year":"2019","unstructured":"von Geldern TW et al (2019) Discovery of ABBV-4083, a novel analog of Tylosin A that has potent anti-Wolbachia and anti-filarial activity. PLoS Negl Trop Dis 13(2):e0007159. https:\/\/doi.org\/10.1371\/journal.pntd.0007159","journal-title":"PLoS Negl Trop Dis"},{"key":"7411_CR26","unstructured":"WHO (2020) Elimination of human onchocerciasis: progress report, 2019\u20132020. Wkly Epidemiol Rec 95:545\u2013556"}],"container-title":["Parasitology Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00436-021-07411-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00436-021-07411-5\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00436-021-07411-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,6]],"date-time":"2022-04-06T07:03:58Z","timestamp":1649228638000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00436-021-07411-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,10]]},"references-count":26,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2022,4]]}},"alternative-id":["7411"],"URL":"https:\/\/doi.org\/10.1007\/s00436-021-07411-5","relation":{},"ISSN":["0932-0113","1432-1955"],"issn-type":[{"value":"0932-0113","type":"print"},{"value":"1432-1955","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,10]]},"assertion":[{"value":"30 August 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 December 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 January 2022","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All participants of the prior studies (DOLF, ISRCTN50035143; MoRiOn, ISRCTN43697583; AWOL-Mino, ISRCTN68861628) gave their informed consent prior to study procedures according to local and national laws and in line with the requirements of Good Clinical Practice. All participants gave their informed consent for their samples to be biobanked and used for biomarker analysis. The informed consent included the consent to participate and consent to publication. The animal work was approved according to the rules of the animal welfare and protection act and national laws, specifically in accordance with the European Union Directive 2010\/63\/EU. Experiments were carried out after approval by the Landesamt f\u00fcrNatur, Umwelt und Verbraucherschutz, Cologne, Germany (AZ 84\u201302.04.2015.A507).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}]}}