Comparison of Three Different DNA Extraction Methods for Linguatula serrata as a Food Born Pathogen
AbstractBackground: One of the most important items in molecular characterization of food-borne pathogens is high quality genomic DNA. In this study, we investigated three protocols and compared their simplicity, duration and costs for extracting genomic DNA from Linguatula serrata.Methods: The larvae were collected from the sheep’s visceral organs from the Yazd Slaughterhouse during May 2013. DNA extraction was done in three different methods, including commercial DNA extraction kit, Phenol Chloroform Isoamylalcohol (PCI), and salting out. Extracted DNA in each method was assessed for quantity and quality using spectrophotometery and agarose gel electrophoresis, respectively.Results: The less duration was regarding to commercial DNA extraction kit and then salting out protocol. The cost benefit one was salting out and then PCI method. The best quantity was regarding to PCI with 72.20±29.20 ng/μl, and purity of OD260/OD280 in 1.76±0.947. Agarose gel electrophoresis for assessing the quality found all the same.Conclusion: Salting out is introduced as the best method for DNA extraction from L. seratta as a food-borne pathogen with the least costand appropriate purity. Although, the best purity was regarding to PCI but PCI is not safe as salting out. In addition, the duration of salting out was less than PCI. The least duration was seen in commercial DNA extraction kit, but it is expensive and therefore is not recommended for developing countries where consumption of offal is common.
Eslami G, Hajimohammadi B, Gholamrezaei M, Khalatbary S, Zohortabar A, Ardian M. Practical approach for DNA extraction of food born Linguatula serrata nymphs: An analytical method. Galen Med J. 2014; 3 (2): 115-19.
Oryan A, Sadjjadi S, Mehrabani D, Rezaei M. The status of Linguatula serrata infection of stray dogs in Shiraz, Iran. Comp Clin Path. 2008;17(1):55-60.
Rezaei H, Ashrafihelan J, Nematollahi A, Mostafavi E. The prevalence of Linguatula serrata nymphs in goats slaughtered in Tabriz, Iran. J Parasit Dis. 2012;36(2):200-2.
Tavassoli M, Tajic H, Dalir-Naghadeh B, Hariri F. Prevalence of Linguatula serrata nymphs and gross changes of infected mesenteric lymph nodes in sheep in Urmia, Iran. Small Rumin Res. 2007;72(1):73-6.
Oryan A, Moghaddar N, Hanifepour MR. Arthropods recovered from the visceral organs of camel with special reference to their incidence and pathogenesis in Fars Province of Iran. Indian J Anim Sci. 1993; 63(3): 290-293.
Haddadzadeh H, Athari SS, Hajimohammadi B. The first record of Linguatula serrata infection of two-humped camel (Camelus bactrinus) in Iran. Iran J Parasitol. 2009;4(1):59-61.
Hajimohammadi B, Akhondzadeh Basti A, Shirali S. Impact of sodium chloride and heat on survival time of Linguatula Serrata nymphs in vitro: An experimental study. J Health Res. 2012;1(1):54-61.
Akhondzadeh Basti A, Haddadzadeh H, Tajik H et al. Effect of different temperature conditions on survival time of Linguatula serrata nymphs. HVM Bioflux. 2011;3(2):76-82.
Bamorovat M, Borhani-Zarandi M, Mostafavi M, Kheirandish R, Sharifi I, Radfar MH. The prevalence of Linguatula serrata nymphs in mesenteric and mediastinal lymph nodes in one-humped camels (Camelus dromedarius) slaughtered in Rafsanjan slaughterhouse, Iran. J Parasit Dis. 2014; 38(4):374-377.
Oryan A, Khordadmehr M, Ranjbar VR. Prevalence, biology, pathology, and public health importance of linguatulosis of camel in Iran. Trop Anim Health Prod. 2011;43(6):1225-31.
Khalil G, Haddad C, Otrock ZK, Jaber F, Farra A. Halzoun, an allergic pharyngitis syndrome in Lebanon: the trematode Dicrocoelium dendriticum as an additional cause. Acta Trop. 2012; 25:115-118.
Riley J. The biology of pentastomids. Adv Parasitol. 1986; 25:45-128.
Almeida De Oliveira, Christoffersen W, Lindsey, M. A cladistic approach to relationships in Pentastomida. J Parasitol. 1999; 85(4):695-704.
Poore GC. The nomenclature of the Recent Pentastomida (Crustacea), with a list of species and available names. Syst Parasitol. 2012; 82 (3): 211-40.
Christoffersen M, De Assis JE. A systematic monograph of the Recent Pentastomida, with a compilation of their hosts. Zool Med Leiden. 2013; 87(1),29: 1-206.
Abele L, Kim W, Felgenhauer B. Molecular evidence for inclusion of the phylum Pentastomida in the Crustacea. Mol Biol Evol. 1989; 6(6):685.
Lavrov DV, Brown WM, Boore JL. Phylogenetic position of the Pentastomida and (pan) crustacean relationships. Proc R Soc Lond B Biol Sci. 2004;271(1538):537-44.
von Reumont BM, Meusemann K, Szucsich NU et al. Can comprehensive background knowledge be incorporated into substitution models to improve phylogenetic analyses? A case study on major arthropod relationships. BMC Evol Biol. 2009;9(1):119.
Sanders KL, Lee MS. Arthropod molecular divergence times and the Cambrian origin of pentastomids. System Biodivers. 2010;8(1):63-74.
Koehsler M, Walochnik J, Georgopoulos M et al. Linguatula serrata tongue worm in human eye, Austria. Emerg Infect Dis. 2011;17(5):870-2.
Tappe D, Meyer M, Oesterlein A et al. Transmission of Armillifer armillatus ova at snake farm, The Gambia, West Africa. Emerg Infect Dis. 2011;17(2):251.
Sambrook J, Russell DW. Transcriptional run-on assays. Cold Spring Harbor Protocols. 2006;(1):doi:10.1101/pdb. prot3956.
Hajimohammadi B, Eslami G, Oryan A, Zohourtabar A, Pourmirzaei TH, Moghaddam AM. Molecular identification of Sarcocystis hominis in native cattle of central Iran: A case report. Trop Biomed. 2014;31(1):183-6.
Boughattas S, Salehi R. Molecular approaches for detection and identification of foodborne pathogens. J Food Qual Hazards Control. 2014; 1 (1) :1-6.
Alzohairy AM. Letter to the editor. J Food Qual Hazards Control. 2014; 1 (1) :35.
Bhaganna P, Volkers RJ, Bell AN, Kluge K, Timson DJ, McGrath JW, et al. Hydrophobic substances induce water stress in microbial cells. Microb Biotechnol. 2010;3(6):701-16.
Eaglestone SS, Ruddock LW, Cox BS, Tuite MF. Guanidine hydrochloride blocks a critical step in the propagation of the prion-like determinant [PSI+] of Saccharomyces cerevisiae. Proc Natl Acad Sci India Sect B Biol Sci. 2000;97(1):240-4.
Freschi CR, Oliveira CJBd. Comparison of DNA-extraction methods and selective enrichment broths on the detection of Salmonella Typhimurium in swine feces by polymerase chain reaction (PCR). Braz J Microbiol. 2005;36(4):363-7.
Asadzadeh N, Javanmard A, Nassiry M. Comparison of rapid DNA extraction techniques for conventional PCR-RFLP analysis from mammalian whole blood cells. J Mol Genet. 2010;2(3):32-5.
Davoudi A, Tarang A, Aleyasin SA, Salehi A, Seighalani R, Tahmoressi F. Evaluation of two DNA extraction methods from maternal plasma for using in non-invasive bovine fetus gender determination. Iran J Reprod Med. 2012;10(6):523-30.
Sambrook J, Russell DW. Purification of nucleic acids by extraction with phenol: chloroform. Cold Spring Harbor Protocols. 2006;(1): doi: 10.1101/pdb. prot4455.
Tan SC, Yiap BC. DNA, RNA, and protein extraction: the past and the present. J Biomed Biotechnol. 2009; (1):1-10.