Abstract
Fascioliasis is a parasitic infection typically caused by two common parasites of class Trematodo, genus Fasciola, namely Fasciola hepatica and Fasciola gigantica. The widespread of these species in water and food makes fascioliasis become a global zoonotic disease that affects 2.4 million people in more than 75 countries worldwide. Typically, F. hepatica and F. gigantica can be recognized by parasitological techniques to detect Fasciola spp. eggs, immunological techniques to detect worm-specific antibodies, or by molecular techniques such as PCR to detect parasitic genomic DNA. Recently, miRNAs have been raised as a key regulator and potential diagnostic biomarkers of diseases, including parasitic infection. An isothermal PCR called loop-mediated isothermal amplification (LAMP) is rapid, sensitive, and its amplification process is so extensive that making LAMP well-suited for field diagnostics. LAMP reactions for miRNA detection have been introduced and were able to detect the target miRNA amounts in the wide range of 1.0 amol to 1.0 pmol, exhibiting high selectivity to differentiate one-base between miRNA sequences. Here, we introduced a modified LAMP to detect a species-specific miRNA of F. hepatica and F. gigantica. Our method did not demand an initial heating step and the reactions had a high sensitivity that greater than 1000 times in comparison to that reported in previous studies. Most importantly, the technique could perform well with parasitic miRNA presenting in bovine serum samples without sophisticated equipment required. These results create a promising technique basis for some novel and simple device to diagnose fascioliasis and other parasitic infection diseases at point-of-care.
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References
Asaga S, Kuo C, Nguyen T et al (2011) Direct serum assay for microRNA-21 concentrations in early and advanced breast cancer. Clin Chem 57:84–91. https://doi.org/10.1373/clinchem.2010.151845
Brase JC, Wuttig D, Kuner R, Sültmann H (2010) Serum microRNAs as non-invasive biomarkers for cancer. Mol Cancer 9:306. https://doi.org/10.1186/1476-4598-9-306
Britton C, Winter AD, Gillan V, Devaney E (2014) microRNAs of parasitic helminths–identification, characterization and potential as drug targets. Int J Parasitol Drugs Drug Resist 4:85–94. https://doi.org/10.1016/j.ijpddr.2014.03.001
Britton C, Winter AD, Marks ND et al (2015) Application of small RNA technology for improved control of parasitic helminths. Vet Parasitol 212:47–53. https://doi.org/10.1016/j.vetpar.2015.06.003
Buck AH, Coakley G, Simbari F et al (2014) Exosomes secreted by nematode parasites transfer small RNAs to mammalian cells and modulate innate immunity. Nat Commun 5:5488. https://doi.org/10.1038/ncomms6488
Buser JR, Diesburg S, Singleton J et al (2015) Precision chemical heating for diagnostic devices. Lab Chip 15:4423–4432. https://doi.org/10.1039/c5lc01053e
Cai P, Gobert GN, You H et al (2015) Circulating miRNAs: potential novel biomarkers for hepatopathology progression and diagnosis of schistosomiasis japonica in two murine models. PLoS Negl Trop Dis 9:e0003965. https://doi.org/10.1371/journal.pntd.0003965
Cai P, Gobert GN, McManus DP (2016) MicroRNAs in parasitic helminthiases: current status and future perspectives. Trends Parasitol 32:71–86. https://doi.org/10.1016/j.pt.2015.09.003
Chen C, Ridzon DA, Broomer AJ et al (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33:e179. https://doi.org/10.1093/nar/gni178
Chim SSC, Shing TKF, Hung ECW et al (2008) Detection and characterization of placental microRNAs in maternal plasma. Clin Chem 54:482–490. https://doi.org/10.1373/clinchem.2007.097972
de Waal T (2016) Diseases of dairy animals: parasites, internal: liver flukes. In: Reference module in food science. Elsevier
Holz A, Streit A (2017) Gain and loss of small RNA classes—characterization of small RNAs in the parasitic nematode family strongyloididae. Genome Biol Evol 9:2826–2843. https://doi.org/10.1093/gbe/evx197
Hong C-Y, Chen X, Liu T et al (2013) Ultrasensitive electrochemical detection of cancer-associated circulating microRNA in serum samples based on DNA concatamers. Biosens Bioelectron 50:132–136. https://doi.org/10.1016/j.bios.2013.06.040
Hoy AM, Lundie RJ, Ivens A et al (2014) Parasite-derived MicroRNAs in host serum as novel biomarkers of helminth infection. PLoS Negl Trop Dis 8:e2701. https://doi.org/10.1371/journal.pntd.0002701
Khan N, Mironov G, Berezovski MV (2016) Direct detection of endogenous MicroRNAs and their post-transcriptional modifications in cancer serum by capillary electrophoresis-mass spectrometry. Anal Bioanal Chem 408:2891–2899. https://doi.org/10.1007/s00216-015-9277-y
Leshkowitz D, Horn-Saban S, Parmet Y, Feldmesser E (2013) Differences in microRNA detection levels are technology and sequence dependent. RNA N Y N 19:527–538. https://doi.org/10.1261/rna.036475.112
Li C, Li Z, Jia H, Yan J (2011) One-step ultrasensitive detection of microRNAs with loop-mediated isothermal amplification (LAMP). Chem Commun 47:2595–2597. https://doi.org/10.1039/C0CC03957H
Manzano-Román R, Siles-Lucas M (2012) MicroRNAs in parasitic diseases: potential for diagnosis and targeting. Mol Biochem Parasitol 186:81–86. https://doi.org/10.1016/j.molbiopara.2012.10.001
Mar-Aguilar F, Trevino V, Salinas-Hernández JE et al (2013) Identification and characterization of microRNAS from entamoeba histolytica HM1-IMSS. PLoS ONE 8:e68202. https://doi.org/10.1371/journal.pone.0068202
Marcilla A, Trelis M, Cortés A et al (2012) Extracellular vesicles from parasitic helminths contain specific excretory/secretory proteins and are internalized in intestinal host cells. PLoS ONE 7:e45974. https://doi.org/10.1371/journal.pone.0045974
Notomi T, Okayama H, Masubuchi H et al (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63. https://doi.org/10.1093/nar/28.12.e63
Patnaik SK, Kannisto ED, Mallick R et al (2017) Whole blood microRNA expression may not be useful for screening non-small cell lung cancer. PLoS ONE 12:e0181926. https://doi.org/10.1371/journal.pone.0181926
Ren Y, Deng H, Shen W, Gao Z (2013) A highly sensitive and selective electrochemical biosensor for direct detection of microRNAs in serum. Anal Chem 85:4784–4789. https://doi.org/10.1021/ac400583e
Schultz NA, Dehlendorff C, Jensen BV et al (2014) MicroRNA biomarkers in whole blood for detection of pancreatic cancer. JAMA 311:392–404. https://doi.org/10.1001/jama.2013.284664
Shah KG, Guelig D, Diesburg S et al (2015) Design of a new type of compact chemical heater for isothermal nucleic acid amplification. PLoS ONE 10:e0139449. https://doi.org/10.1371/journal.pone.0139449
Singleton J, Zentner C, Buser J et al (2013) Instrument-free exothermic heating with phase change temperature control for paper microfluidic devices. Proc SPIE Int Soc Opt Eng 8615:86150. https://doi.org/10.1117/12.2005928
Singleton J, Osborn JL, Lillis L et al (2014) Electricity-free amplification and detection for molecular point-of-care diagnosis of HIV-1. PLoS ONE 9:e113693. https://doi.org/10.1371/journal.pone.0113693
Tanner NA, Zhang Y, Evans TC (2015) Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques 58:59–68. https://doi.org/10.2144/000114253
Tian T, Wang J, Zhou X (2015) A review: microRNA detection methods. Org Biomol Chem 13:2226–2238. https://doi.org/10.1039/C4OB02104E
Tolan RW (2011) Fascioliasis due to Fasciola hepatica and Fasciola gigantica infection: an update on this ‘Neglected’ neglected tropical disease. Lab Med 42:107–116. https://doi.org/10.1309/LMLFBB8PW4SA0YJI
Tritten L, Burkman E, Moorhead A et al (2014) Detection of circulating parasite-derived MicroRNAs in filarial infections. PLoS Negl Trop Dis 8:e2971. https://doi.org/10.1371/journal.pntd.0002971
Válóczi A, Hornyik C, Varga N et al (2004) Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. Nucleic Acids Res 32:e175. https://doi.org/10.1093/nar/gnh171
Wang K, Yuan Y, Cho J-H et al (2012) Comparing the MicroRNA spectrum between serum and plasma. PLoS ONE 7:e41561. https://doi.org/10.1371/journal.pone.0041561
Xin L, Gao J, Wang D et al (2017) Novel blood-based microRNA biomarker panel for early diagnosis of chronic pancreatitis. Sci Rep 7:40019. https://doi.org/10.1038/srep40019
Xu M-J, Ai L, Fu J-H et al (2012) Comparative characterization of MicroRNAs from the liver flukes Fasciola gigantica and F. hepatica. PLoS ONE 7:e53387. https://doi.org/10.1371/journal.pone.0053387
Yyusnita Y, Norsiah MD, Zakiah I et al (2012) MicroRNA (miRNA) expression profiling of peripheral blood samples in multiple myeloma patients using microarray. Malays J Pathol 34:133–143
Zamanian M, Fraser LM, Agbedanu PN et al (2015) Release of small RNA-containing exosome-like vesicles from the human filarial parasite Brugia malayi. PLoS Negl Trop Dis 9:e0004069. https://doi.org/10.1371/journal.pntd.0004069
Zheng Y, Cai X, Bradley JE (2013) microRNAs in parasites and parasite infection. RNA Biol 10:371–379. https://doi.org/10.4161/rna.23716
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This research is funded by NTTU Foundation for Science and Technology Development under Grant Number 2018.01.13.
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Tran, D.H., Phung, H.T.T. Detecting Fasciola hepatica and Fasciola gigantica microRNAs with loop-mediated isothermal amplification (LAMP). J Parasit Dis 44, 364–373 (2020). https://doi.org/10.1007/s12639-019-01164-w
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DOI: https://doi.org/10.1007/s12639-019-01164-w