Natalia Souza DE De Godoy, Vera Demarchi Aiello, Regina Maia de Souza, Thelma Okay, Lucia Maria Almeida Braz
Laboratory of Investigation in Medical Parasitology, Clínicas Hospital, Faculty of Medicine, University of São Paulo, São Paulo, Brazil.
Dept. of Infectious and Parasitic Diseases, Clínicas Hospital, Faculty of Medicine, University of São Paulo, São Paulo, Brazil.
Iran J Parasitol. 2018 Oct-Dec;13(4):655-660.
Patients coinfected with /HIV can develop atypical forms of visceral leishmaniasis (VL), making it indispensable to identify the etiological agent. We are presenting a post-mortem specie definition by ITS1-PCR-RFLP in a larynx tissue of a patient presented coinfection /HIV. This patient was from a leishmaniasis endemic region in São Paulo (SP), Brazil, and was diagnosed clinically with mucocutaneous leishmaniasis. Before a rK39 immunochromatographic test positive, a tiny stored paraffin-embedded larynx tissue was obtained post-mortem and submitted to 3 conventional PCR assays: kDNA (K20/K22 and RV1/RV2), and ITS1 (LITSR/L5.8S). The last one was followed by RFLP (III) and analyzed by 4% Metaphor agarose gel electrophoresis. genus and () subgenus were defined by kDNA-PCR, with K20/K22 (120 bp) and RV1/RV2 (145 bp), respectively. ITS1-PCR-RFLP identified species visualized by the restriction patterns of 180, 70 and 50 bp. This case draws attention to the necessity for a clear identification of the etiological agent causing infection, especially in endemic regions of cutaneous and visceral leishmaniasis, and particularly in patients with comorbidities who often present atypical forms of the disease. () , which is usually responsible for VL, had changed its clinical spectrum for mucocutaneous. Unequivocal identification was carried out by ITS-PCR-RFLP, therefore confirming rK39 result. These techniques, which complemented each other, have a convenient cost-benefit ratio that makes them suitable to be applied in developing countries.
合并感染艾滋病病毒（HIV）的患者可能会出现非典型形式的内脏利什曼病（VL），因此确定病原体至关重要。我们通过ITS1-PCR-RFLP对一名合并感染HIV的患者喉部组织进行了尸检物种鉴定。该患者来自巴西圣保罗（SP）的利什曼病流行地区，临床诊断为皮肤黏膜利什曼病。在rK39免疫层析试验呈阳性之前，患者死后获取了一小块保存的石蜡包埋喉部组织，并进行了3种常规PCR检测：kDNA（K20/K22和RV1/RV2）以及ITS1（LITSR/L5.8S）。最后一项检测之后进行RFLP（III），并通过4%的Metaphor琼脂糖凝胶电泳进行分析。通过kDNA-PCR分别用K20/K22（120 bp）和RV1/RV2（145 bp）确定了属和（）亚属。ITS1-PCR-RFLP鉴定出了通过180、70和50 bp的限制性图谱可见的物种。该病例提醒人们，明确鉴定引起感染的病原体非常必要，尤其是在皮肤和内脏利什曼病的流行地区，特别是在经常出现非典型疾病形式的合并症患者中。通常导致VL的（），其临床谱已转变为皮肤黏膜型。通过ITS-PCR-RFLP进行了明确鉴定，从而证实了rK39的结果。这些相互补充的技术具有便利的成本效益比，使其适合在发展中国家应用。
