Tolba Mohammed Essa Marghany, Huseein Enas Abdelhameed Mahmoud, Farrag Haiam Mohamed Mahmoud, Mohamed Hanan El Deek, Kobayashi Seiki, Suzuki Jun, Ali Tarek Ahmed Mohamed, Sugano Sumio
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.
PLoS Negl Trop Dis. 2016 Jul 14;10(7):e0004841. doi: 10.1371/journal.pntd.0004841. eCollection 2016 Jul.
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.
自由生活阿米巴在全球范围内均有存在。它们能够在不同环境中生存,在某些情况下会引发人类疾病。棘阿米巴属以导致人类视力威胁性角膜炎而闻名。自由生活阿米巴角膜炎在发展中国家更为常见。瓦尔坎普夫科的阿米巴很少被报道会引发此类病症。最近从洞穴中鉴定出一个新属——洞穴异瓦尔坎普夫阿米巴,但尚无关于其对人类致病性的数据。我们试图运用形态学和分子技术,在一名埃及角膜炎患者中鉴定出致病的自由生活阿米巴。
使用单菌培养系统培养致病性阿米巴。通过形态学特征以及18S核糖体RNA亚基DNA扩增和测序进行鉴定。对实验用兔子评估其致病性以及诱发角膜炎的能力。
洞穴异瓦尔坎普夫阿米巴被鉴定为人类角膜炎的病因。对18S核糖体亚基DNA的全序列进行了测序和组装。埃及菌株与在斯洛文尼亚分离出的SK1菌株密切相关。利用动物模型证实了其诱发角膜炎的能力。
这是首次报道洞穴异瓦尔坎普夫阿米巴作为人类病原体。结合分子和形态学鉴定对于正确诊断导致人类角膜炎的阿米巴至关重要。需要使用不同的引物对并对扩增的DNA进行测序以防止误诊。