Jundishapur Journal of Microbiology

Published by: Kowsar

Evaluation of Three Protocols of DNA Extraction for Detection of Giardia duodenalis in Human Fecal Specimens

Fatemeh Asgarian 1 , 2 , Mehdi Tavalla 1 , 2 , * , Ali Teimoori 3 , Nozhat Zebardast 4 and Bahman Cheraghian 5
Authors Information
1 Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
2 Department of Parasitology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
3 Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
4 Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran
5 Research Center for Infectious Diseases of Digestive System, Department of Biostatistics and Epidemiology, School of Public Health, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Article information
  • Jundishapur Journal of Microbiology: April 2018, 11 (4); e63096
  • Published Online: March 10, 2018
  • Article Type: Research Article
  • Received: October 21, 2017
  • Revised: February 10, 2018
  • Accepted: February 17, 2018
  • DOI: 10.5812/jjm.63096

To Cite: Asgarian F, Tavalla M, Teimoori A, Zebardast N, Cheraghian B. Evaluation of Three Protocols of DNA Extraction for Detection of Giardia duodenalis in Human Fecal Specimens, Jundishapur J Microbiol. 2018 ; 11(4):e63096. doi: 10.5812/jjm.63096.

Copyright © 2018, Jundishapur Journal of Microbiology. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited
1. Background
2. Objectives
3. Methods
4. Results
5. Discussion
6. Conclusions
  • 1. Aguiar JM, Silva SO, Santos VA, Rossa GA, Taniwaki SA, Gregori F, et al. Multilocus amplification of genomic DNA from single cysts of Giardia duodenalis separated using micromanipulation technique. Exp Parasitol. 2015;157:84-7. doi: 10.1016/j.exppara.2015.07.004. [PubMed: 26172406].
  • 2. Koloren Z, Seferoglu O, Karanis P. Occurency of Giardia duodenalis assemblages in river water sources of Black Sea, Turkey. Acta Trop. 2016;164:337-44. doi: 10.1016/j.actatropica.2016.09.025. [PubMed: 27697482].
  • 3. Koehler AV, Jex AR, Haydon SR, Stevens MA, Gasser RB. Giardia/giardiasis - a perspective on diagnostic and analytical tools. Biotechnol Adv. 2014;32(2):280-9. doi: 10.1016/j.biotechadv.2013.10.009. [PubMed: 24189092].
  • 4. Babaei Z, Oormazdi H, Rezaie S, Rezaeian M, Razmjou E. Giardia intestinalis: DNA extraction approaches to improve PCR results. Exp Parasitol. 2011;128(2):159-62. doi: 10.1016/j.exppara.2011.02.001. [PubMed: 21315715].
  • 5. Uda-Shimoda CF, Colli CM, Pavanelli MF, Falavigna-Guilherme AL, Gomes ML. Simplified protocol for DNA extraction and amplification of 2 molecular markers to detect and type Giardia duodenalis. Diagn Microbiol Infect Dis. 2014;78(1):53-8. doi: 10.1016/j.diagmicrobio.2013.09.008. [PubMed: 24207076].
  • 6. Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev. 2011;24(1):110-40. doi: 10.1128/CMR.00033-10. [PubMed: 21233509].
  • 7. Jerez Puebla LE, Nunez FA, Martinez Silva I, Rojas Rivero L, Martinez Gonzalez M, Mendez Sutil Y, et al. Molecular Characterization and Risk Factors of Giardia duodenalis among School Children from La Habana, Cuba. J Parasitol Res. 2015;2015:378643. doi: 10.1155/2015/378643. [PubMed: 26693345].
  • 8. Paulos S, Mateo M, de Lucio A, Hernandez-de Mingo M, Bailo B, Saugar JM, et al. Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp. J Microbiol Methods. 2016;127:68-73. doi: 10.1016/j.mimet.2016.05.020. [PubMed: 27241828].
  • 9. de Lucio A, Martinez-Ruiz R, Merino FJ, Bailo B, Aguilera M, Fuentes I, et al. Molecular Genotyping of Giardia duodenalis Isolates from Symptomatic Individuals Attending Two Major Public Hospitals in Madrid, Spain. PLoS One. 2015;10(12). e0143981. doi: 10.1371/journal.pone.0143981. [PubMed: 26641082].
  • 10. Bartelt LA, Attias E, Black J. Epidemiology of cryptosporidiosis and giardiasis: What pediatricians need to know. Curr Trop Med Rep. 2016;3(3):108-14. doi: 10.1007/s40475-016-0081-3.
  • 11. Halliez MC, Buret AG. Extra-intestinal and long term consequences of Giardia duodenalis infections. World J Gastroenterol. 2013;19(47):8974-85. doi: 10.3748/wjg.v19.i47.8974. [PubMed: 24379622].
  • 12. Al-Mekhlafi HM, Al-Maktari MT, Jani R, Ahmed A, Anuar TS, Moktar N, et al. Burden of Giardia duodenalis infection and its adverse effects on growth of schoolchildren in rural Malaysia. PLoS Negl Trop Dis. 2013;7(10). e2516. doi: 10.1371/journal.pntd.0002516. [PubMed: 24205426].
  • 13. Fletcher SM, Stark D, Harkness J, Ellis J. Enteric protozoa in the developed world: a public health perspective. Clin Microbiol Rev. 2012;25(3):420-49. doi: 10.1128/CMR.05038-11. [PubMed: 22763633].
  • 14. Kuk S, Yazar S, Cetinkaya U. Stool sample storage conditions for the preservation of Giardia intestinalis DNA. Mem Inst Oswaldo Cruz. 2012;107(8):965-8. doi: 10.1590/S0074-02762012000800001. [PubMed: 23295744].
  • 15. Asher AJ, Waldron LS, Power ML. Evaluation of a PCR protocol for sensitive detection of Giardia intestinalis in human faeces. Parasitol Res. 2012;110(2):853-8. doi: 10.1007/s00436-011-2565-3. [PubMed: 21800123].
  • 16. Zebardast N, Yeganeh F, Gharavi MJ, Abadi A, Seyyed Tabaei SJ, Haghighi A. Simultaneous detection and differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii, Giardia lamblia and Cryptosporidium spp. in human fecal samples using multiplex PCR and qPCR-MCA. Acta Trop. 2016;162:233-8. doi: 10.1016/j.actatropica.2016.07.004. [PubMed: 27423396].
  • 17. Eligio-Garcia L, Adrian CC, Apolinar CE, Enedina JC. Assemblages of Giardia duodenalis Isolates from Dogs by Amplification and Restriction of tpi and β-Giardin Genes and Sequencing of SSU rDNA Gene. Open J Vet Med. 2013;3(3):222-7. doi: 10.4236/ojvm.2013.33035.
  • 18. Geurden T, Vercruysse J, Claerebout E. Is Giardia a significant pathogen in production animals? Exp Parasitol. 2010;124(1):98-106. doi: 10.1016/j.exppara.2009.03.001. [PubMed: 19285075].
  • 19. Traversa D, Otranto D, Milillo P, Latrofa MS, Giangaspero A, Di Cesare A, et al. Giardia duodenalis sub-Assemblage of animal and human origin in horses. Infect Genet Evol. 2012;12(8):1642-6. doi: 10.1016/j.meegid.2012.06.014. [PubMed: 22771626].
  • 20. Huey CS, Mahdy MA, Al-Mekhlafi HM, Nasr NA, Lim YA, Mahmud R, et al. Multilocus genotyping of Giardia duodenalis in Malaysia. Infect Genet Evol. 2013;17:269-76. doi: 10.1016/j.meegid.2013.04.013. [PubMed: 23624189].
  • 21. Schrader C, Schielke A, Ellerbroek L, Johne R. PCR inhibitors - occurrence, properties and removal. J Appl Microbiol. 2012;113(5):1014-26. doi: 10.1111/j.1365-2672.2012.05384.x. [PubMed: 22747964].
  • 22. Sepahvand A, Pestehchian N, Yousefi HA, Gharehbaba RP. Comparison and evaluation of four methods for extracting DNA from Giardia duodenalis cysts for PCR targeting the tpi gene. J Parasit Dis. 2017;41(1):263-7. doi: 10.1007/s12639-016-0790-5. [PubMed: 28316423].
  • 23. Pestechian N, Rasekh H, Rostami-Nejad M, Yousofi HA, Hosseini-Safa A. Molecular identification of Giardia lamblia; is there any correlation between diarrhea and genotyping in Iranian population? Gastroenterol Hepatol Bed Bench. 2014;7(3):168-72. [PubMed: 25120898].
  • 24. Ryan U, Caccio SM. Zoonotic potential of Giardia. Int J Parasitol. 2013;43(12-13):943-56. doi: 10.1016/j.ijpara.2013.06.001. [PubMed: 23856595].
  • 25. Minetti C, Lamden K, Durband C, Cheesbrough J, Fox A, Wastling JM. Determination of Giardia duodenalis assemblages and multi-locus genotypes in patients with sporadic giardiasis from England. Parasit Vectors. 2015;8:444. doi: 10.1186/s13071-015-1059-z. [PubMed: 26338670].
  • 26. Heyworth MF. Giardia duodenalis genetic assemblages and hosts. Parasite. 2016;23:13. doi: 10.1051/parasite/2016013. [PubMed: 26984116].
  • 27. Ramirez JD, Heredia RD, Hernandez C, Leon CM, Moncada LI, Reyes P, et al. Molecular diagnosis and genotype analysis of Giardia duodenalis in asymptomatic children from a rural area in central Colombia. Infect Genet Evol. 2015;32:208-13. doi: 10.1016/j.meegid.2015.03.015. [PubMed: 25795384].
  • 28. Rayani M, Zasmy Unyah N, Hatam G. Molecular Identification of Giardia duodenalis Isolates from Fars Province, Iran. Iran J Parasitol. 2014;9(1):70-8. [PubMed: 25642262].
  • 29. Elwin K, Fairclough HV, Hadfield SJ, Chalmers RM. Giardia duodenalis typing from stools: a comparison of three approaches to extracting DNA, and validation of a probe-based real-time PCR typing assay. J Med Microbiol. 2014;63(Pt 1):38-44. doi: 10.1099/jmm.0.066050-0. [PubMed: 24085818].
  • 30. Nantavisai K, Mungthin M, Tan-ariya P, Rangsin R, Naaglor T, Leelayoova S. Evaluation of the sensitivities of DNA extraction and PCR methods for detection of Giardia duodenalis in stool specimens. J Clin Microbiol. 2007;45(2):581-3. doi: 10.1128/JCM.01823-06. [PubMed: 17122010].
  • 31. Jiang J, Alderisio KA, Singh A, Xiao L. Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates and for relief of PCR inhibitors. Appl Environ Microbiol. 2005;71(3):1135-41. doi: 10.1128/AEM.71.3.1135-1141.2005. [PubMed: 15746310].
  • 32. Hawash Y. DNA extraction from protozoan oocysts/cysts in feces for diagnostic PCR. Korean J Parasitol. 2014;52(3):263-71. doi: 10.3347/kjp.2014.52.3.263. [PubMed: 25031466].
  • 33. Adamska M, Leonska-Duniec A, Maciejewska A, Sawczuk M, Skotarczak B. Comparison of efficiency of various DNA extraction methods from cysts of Giardia intestinalis measured by PCR and TaqMan real time PCR. Parasite. 2010;17(4):299-305. doi: 10.1051/parasite/2010174299. [PubMed: 21275235].
  • 34. Psifidi A, Dovas CI, Bramis G, Lazou T, Russel CL, Arsenos G, et al. Comparison of eleven methods for genomic DNA extraction suitable for large-scale whole-genome genotyping and long-term DNA banking using blood samples. PLoS One. 2015;10(1). e0115960. doi: 10.1371/journal.pone.0115960. [PubMed: 25635817].
  • 35. Abbaszadegan MR, Velayati A, Tavasoli A, Dadkhah E. Rapid DNA extraction protocol from stool, suitable for molecular genetic diagnosis of colon cancer. Iran Biomed J. 2007;11(3):203-8. [PubMed: 18051782].
  • 36. Buckingham L, Flaws ML. Molecular Diagnostics: Fundamentals, Methods, & Clinical Applications. F.A. Davis; 2007.
  • 37. Rai AK, Chakravorty R, Paul J. Detection of Giardia, Entamoeba, and Cryptosporidium in unprocessed food items from northern India. World Journal of Microbiology and Biotechnology. 2008;24(12):2879-87. doi: 10.1007/s11274-008-9824-1.
  • 38. Yu X, Van Dyke MI, Portt A, Huck PM. Development of a direct DNA extraction protocol for real-time PCR detection of Giardia lamblia from surface water. Ecotoxicology. 2009;18(6):661-8. doi: 10.1007/s10646-009-0347-9. [PubMed: 19499328].
  • 39. Ghatak S, Muthukumaran RB, Nachimuthu SK. A simple method of genomic DNA extraction from human samples for PCR-RFLP analysis. J Biomol Tech. 2013;24(4):224-31. doi: 10.7171/jbt.13-2404-001. [PubMed: 24294115].
  • 40. Pachner AR, Delaney E. The polymerase chain reaction in the diagnosis of Lyme neuroborreliosis. Ann Neurol. 1993;34(4):544-50. doi: 10.1002/ana.410340407. [PubMed: 8215242].

Featured Image:

Creative Commons License Except where otherwise noted, this work is licensed under Creative Commons Attribution Non Commercial 4.0 International License .

Search Relations:



Create Citiation Alert
via Google Reader

Readers' Comments