Original Article

Molecular Characterization and Analysis of 16S Ribosomal DNA in some Isolates of Demodex folliculorum

Abstract

Background: Demodicosis is one of the most prevalent skin diseases resulting from infestation by Demodex mites. This parasite usually inhabits in follicular infundibulum or sebaceous duct transmitted through close contact with an infested host.

Methods: This study was carried from September 2014 to January 2016 at Tehran University of Medical Sciences, Tehran, Iran. DNA extraction and amplification of 16S ribosomal RNA was performed on four isolates, obtained from four patients and identified morphologically through clearing with 10% Potassium hydroxide (KOH) and microscopical examination. Amplified fragments from the isolates were compared with GenBank database and phylogenetic analysis was carried out using MEGA6 software.

Results: A 390 bp fragment of 16S rDNA was obtained in all isolates and analysis of generated sequences showed high similarity with those submitted to GenBank, previously. Intra-species similarity and distance also showed 99.983% and 0.017, respectively, for the studied isolates. Multiple alignments of the isolates showed Single Nucleotide Polymorphisms (SNPs) in 16S rRNA fragment. Phylogenetic analysis revealed that all 4 isolates clustered with other D. folliculorum, recovered from GenBank database. Our accession numbers KF875587 and KF875589 showed more similarity together in comparison with two other studied isolates.

Conclusion: Mitochondrial 16S rDNA is one of the most suitable molecular barcodes for identification D. folliculorum and this fragment can use for intra-species characterization of the most human-infected mites.

Akbulatova LKh. [The pathogenic role of the mite Demodex and the clinical forms of demodicosis in man]. Vestn Dermatol Venerol. 1966; 40(12):57-61.

Rufli T, Mumcuoglu Y. The hair follicle mites Demodex folliculorum and Demodex brevis: biology and medical importance. A review. Dermatologica. 1981; 162(1):1-11.

Basta-Juzbasic A, Subic JS, Ljubojevic S. Demodex folliculorum in development of dermatitis rosaceiformis steroidica and rosacea-related diseases. Clin Dermatol. 2002; 20(2):135-40.

Baima B, Sticherling M. Demodicidosis revisited. Acta Derm Venereol. 2002; 82(1):3-6.

Aylesworth R, Vance JC. Demodex folliculorum and Demodex brevis in cutaneous biopsies. J Am Acad Dermatol. 1982; 7(5):583-9.

Ozdemir MH, Aksoy U, Sonmez E et al. Prevalence of Demodex in health personnel working in the autopsy room. Am J Forensic Med Pathol. 2005; 26(1):18-23.

Andrews JR. The prevalence of hair follicle mites in caucasian New Zealanders. N Z Med J. 1982; 95(711):451-3.

Orru A, Pintus ML, Pinetti P. Studio sulla infestazione da Demodex folliculorum della cute umana normale. Rass Med Sarda. 1972; 75: 231-44.

Zari J, Abdolmajid F, Masood M et al. Evaluation of the relationship between androgenetic alopecia and demodex infestation. Ind J Dermatol. 2008; 53(2): 64-7.

Moravvej H, Dehghan-Mangabadi M, Abbasian MR et al. Association of rosacea with demodicosis. Arch Iran Med. 2007; 10(2): 199-203.

Talghini S, Fouladi DF, Babaeinejad S et al. Demodex Mite, Rosacea and Skin Melanoma; Coincidence or Association?. Turk Parazitol Derg. 2015; 39(1): 41-6.

Tehrani S, Tizmaghz A, Shabestanipour G. The Demodex mites and their relation with seborrheic and atopic Dermatitis. Asian Pac J Trop Med. 2014; 7S1: S82-4.

Elston DM. Demodex mites as a cause of human disease. Cutis. 2005; 76(5): 294-6.

Santarem VA, Farias MR, Tostes RA. Demodectic mange in fattening pigs in Sao Paulo, Brazil. Vet Parasitol. 2005;131(1-2):169-71.

Heacock CE. Clinical manifestations of demodicosis. J Am Optom Assoc. 1986; 57(12): 914-9.

Elston DM. Demodex mites: facts and controversies. Clin Dermatol. 2010; 28(5): 502-4.

Zhao YE, Cheng J, Hu L et al. Molecular identification and phylogenetic study of Demodex caprae. Parasitol Res. 2014; 113(10): 3601-8.

Zhao YE, Hu L, Ma JX. Molecular identification of four phenotypes of human Demodex mites (Acari: Demodicidae) based on mitochondrial 16S rDNA. Parasitol Res. 2013; 112(11): 3703-11.

Zhao YE, Xu JR, Hu L et al. Complete sequence analysis of 18S rDNA based on genomic DNA extraction from individual Demodex mites (Acari: Demodicidae). Exp Parasitol. 2012; 131(1): 45-51.

Hu L, Zhao YE, Cheng J et al. Molecular identification of four phenotypes of human Demodex in China. Exp Parasitol. 2014; 142: 38-42.

Zhao YE, Wu LP. Phylogenetic relationships in Demodex mites (Acari: Demodicidae) based on mitochondrial 16S rDNA partial sequences. Parasitol Res. 2012; 111(3): 1113-21.

Zhao YE, Wu LP, Hu L et al. Sequencing for complete rDNA sequences (18S, ITS1, 5.8S, ITS2, and 28S rDNA) of Demodex and phylogenetic analysis of Acari based on 18S and 28S rDNA. Parasitol Res. 2012; 111(5): 2109-14.

Zhao YE, Cheng H, Xun M, Wu LP. Extraction and random primer PCR detection of genomic DNA of parasitic mites Demodex folliculorum and Demodex brevis (Acari: Demodicidae). Act Entomol Sin. 2009; 52: 929-33.

Tamura K, Stecher G, Peterson D et al. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 2013; 30(12):2725-9.

Czepita D, Kuzna-Grygiel W, Kosik-Bogacka D. [Investigations on the occurrence as well as the role of Demodex follicuforum and Demodex brevis in the pathogensis of blepharitis]. Klin Oczna. 2005; 107(1-3): 80-2.

Zhao YE, Hu L, Ma JX. Phylogenetic analysis of Demodex caprae based on mitochondrial 16S rDNA sequence. Parasitol Res. 2013; 112(11): 3969-77.

Lv J, Wu S, Zhang Y et al. Assessment of four DNA fragments (COI, 16S rDNA, ITS2, 12S rDNA) for species identification of the Ixodida (Acari: Ixodida). Parasite Vectors. 2014; 7: 93.

Lv J, Wu S, Zhang Y et al. Development of a DNA barcoding system for the Ixodida (Acari: Ixodida). Mitochondrial DNA. 2014; 25(2): 142-9.

De Rojas M, Riazzo C, Callejon R et al. Morphobiometrical and molecular study of two populations of Demodex folliculorum from humans. Parasitol Res. 2012;110(1):227-33.

Sastre N, Ravera I, Villanueva S et al. Phylogenetic relationships in three species of canine Demodex mite based on partial sequences of mitochondrial 16S rDNA. Vet Dermatol. 2012;23(6):509-e101.

Bourdeau P. Variation in size in Demodex canis: from the shortest to the longest forms. Vet Dermatol. 2009;21:213.

Files
IssueVol 12 No 2 (2017) QRcode
SectionOriginal Article(s)
Keywords
Iran Demodex folliculorum 16S rDNA Phylogenetic analysis

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
DANESHPARVAR A, MOWLAVI G, MIRJALALI H, HAJJARAN H, MOBEDI I, NADDAF SR, SHIDFAR M, SADAT MAKKI M. Molecular Characterization and Analysis of 16S Ribosomal DNA in some Isolates of Demodex folliculorum. Iran J Parasitol. 2017;12(2):224-229.