title: Plasma-derived parasitic microRNAs have insufficient concentrations to be used as diagnostic biomarker for detection of Onchocerca volvulus infection or treatment monitoring using LNA-based RT-qPCR.
abstract: River blindness, caused by infection with the filarial nematode Onchocerca volvulus, is a neglected tropical disease affecting millions of people. There is a clear need for diagnostic tools capable of identifying infected patients, but that can also be used for monitoring disease progression and treatment efficacy. Plasma-derived parasitic microRNAs have been suggested as potential candidates for such diagnostic tools. We have investigated whether these parasitic microRNAs are present in sufficient quantity in plasma of Onchocerca-infected patients to be used as a diagnostic biomarker for detection of O. volvulus infection or treatment monitoring. Plasma samples were collected from different sources (23 nodule-positive individuals and 20 microfilaridermic individuals), microRNAs (miRNAs) were extracted using Qiagen miRNeasy kit, and a set of 17 parasitic miRNAs was evaluated on these miRNA extracts using miRCURY Locked Nucleic Acid (LNA) Universal RT microRNA PCR system. Of the 17 miRNAs evaluated, only 7 miRNAs were found to show detectable signal in a number of samples: bma-miR-236-1, bma-miR-71, ov-miR71-22nt, ov-miR-71-23nt, ov-miR-100d, ov-bantam-a, and ov-miR-87-3p. Subsequent melting curve analysis, however, indicated that the signals observed for ov-miR-71 variants and ov-miR-87-3p are non-specific. The other miRNAs only showed positive signal in one or few samples with Cq values just below the cutoff. Our data indicate that parasitic miRNAs are not present in circulation at a sufficiently high level to be used as biomarker for O. volvulus infection or treatment monitoring using LNA-based RT-qPCR analysis.
authors.affiliation: Janssen Diagnostics, Janssen R&D, Turnhoutseweg 30, 2340, Beerse, Belgium. olagatie@its.jnj.com.
authors.forename: Ole
authors.initials: O
authors.lastname: Lagatie
authors.affiliation: Kumasi Centre for Collaborative Research, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana.
authors.forename: Linda
authors.initials: L
authors.lastname: Batsa Debrah
authors.affiliation: Faculty of Allied Health Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana.
authors.forename: Alex
authors.initials: A
authors.lastname: Debrah
authors.affiliation: Janssen Diagnostics, Janssen R&D, Turnhoutseweg 30, 2340, Beerse, Belgium.
authors.forename: Lieven J
authors.initials: LJ
authors.lastname: Stuyver
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000871:Anthelmintics / Q000627:therapeutic use; D015415:Biomarkers / Q000097:blood; D018509:DNA, Helminth / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009841:Oligonucleotides / Q000235:genetics* / Q000378:metabolism; D017181:Onchocerca volvulus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015827:Onchocerciasis, Ocular / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D016896:Treatment Outcome; D055815:Young Adult
chemical_list: D000871:Anthelmintics; D015415:Biomarkers; D018509:DNA, Helminth; D035683:MicroRNAs; D009841:Oligonucleotides; C477371:locked nucleic acid
keywords: Biomarker; Diagnostic; Onchocerca volvulus; Onchocerciasis; River blindness; miRNA