title: Allovahlkampfia spelaea Causing Keratitis in Humans.
abstract: Free-living amoebae are present worldwide. They can survive in different environment causing human diseases in some instances. Acanthamoeba sp. is known for causing sight-threatening keratitis in humans. Free-living amoeba keratitis is more common in developing countries. Amoebae of family Vahlkampfiidae are rarely reported to cause such affections. A new genus, Allovahlkampfia spelaea was recently identified from caves with no data about pathogenicity in humans. We tried to identify the causative free-living amoeba in a case of keratitis in an Egyptian patient using morphological and molecular techniques.
  
  
  
  Pathogenic amoebae were culture using monoxenic culture system. Identification through morphological features and 18S ribosomal RNA subunit DNA amplification and sequencing was done. Pathogenicity to laboratory rabbits and ability to produce keratitis were assessed experimentally.
  
  
  
  Allovahlkampfia spelaea was identified as a cause of human keratitis. Whole sequence of 18S ribosomal subunit DNA was sequenced and assembled. The Egyptian strain was closely related to SK1 strain isolated in Slovenia. The ability to induce keratitis was confirmed using animal model.
  
  
  
  This the first time to report Allovahlkampfia spelaea as a human pathogen. Combining both molecular and morphological identification is critical to correctly diagnose amoebae causing keratitis in humans. Use of different pairs of primers and sequencing amplified DNA is needed to prevent misdiagnosis.
authors.affiliation: Department of Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.|Department of Medical Genomics, Graduate School of Frontier Sciences, The University of Tokyo, Minato-ku, Tokyo, Japan.
authors.forename: Mohammed Essa Marghany
authors.initials: ME
authors.lastname: Tolba
authors.affiliation: Department of Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.
authors.forename: Enas Abdelhameed Mahmoud
authors.initials: EA
authors.lastname: Huseein
authors.affiliation: Department of Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.|Faculty of Applied Medical Science, Shaqra University, Shaqra, Saudi Arabia.
authors.forename: Haiam Mohamed Mahmoud
authors.initials: HM
authors.lastname: Farrag
authors.affiliation: Department of Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.
authors.forename: Hanan El Deek
authors.initials: Hel D
authors.lastname: Mohamed
authors.affiliation: Department of Infectious Diseases, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Seiki
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Division of Clinical Microbiology, Department of Microbiology, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Suzuki
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, Assiut University, Assiut, Egypt.
authors.forename: Tarek Ahmed Mohamed
authors.initials: TA
authors.lastname: Ali
authors.affiliation: Department of Medical Genomics, Graduate School of Frontier Sciences, The University of Tokyo, Minato-ku, Tokyo, Japan.
authors.forename: Sumio
authors.initials: S
authors.lastname: Sugano
mesh_terms: D000048:Acanthamoeba / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D000562:Amebiasis / Q000469:parasitology*; D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004195:Disease Models, Animal; D006801:Humans; D007634:Keratitis / Q000469:parasitology*; D010802:Phylogeny; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D011817:Rabbits
chemical_list: D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S