Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom.
Instituto Nacional de Medicina Tropical (INMeT), ANLIS, Ministerio de Salud de la Nación, Puerto Iguazú, Misiones, Argentina.
PLoS Negl Trop Dis. 2021 Jul 19;15(7):e0009552. doi: 10.1371/journal.pntd.0009552. eCollection 2021 Jul.
Visceral leishmaniasis (VL) is a zoonotic protozoal vector-borne disease that is a major public health challenge. In Argentina, canine (CVL) and human visceral leishmaniasis (HVL) have recently emerged. There is a lack of standardised diagnostic tests for CVL, which hinders control of CVL and HVL.
METHODOLOGY/PRINCIPAL FINDINGS: Sampling was carried out in Puerto Iguazú, Argentina, comprising 190 asymptomatic, oligosymptomatic and polysymptomatic dogs. The following diagnostics were applied: microscopy of lymph node aspirate (LNA); three immunochromatographic rapid diagnostic tests (RDTs), prototype rK28-ICT, rK39-ICT (both Coris BioConcept), commercial rK39 (InBios); ELISA for IgG, IgG1 and IgG2, against rK28, rK39 or crude lysate antigen. DNA detection and analysis, with 30 dogs, was of the ITS1 region using skin samples, and loop-mediated isothermal amplification (LAMP; Eiken Loopamp) of buffy coat, skin scrape or LNA. 15.4% of dogs were positive by LNA microscopy. The rK28 RDT had higher seropositivity rate (61%) than either a prototype rK39 RDT (31.4%) or commercial rK39 RDT (18.8%), without cross-reactivity with six other pathogens. IgG anti-rK39 ELISA antibody titres, but not IgG2, were positively correlated with number of clinical signs. LAMP with LNA had a higher positivity rate than PCR; buffy coat sampling was more sensitive than skin scrape. ITS1 confirmed Leishmania (Leishmania) infantum as the agent of CVL. Leishmania (Viannia) spp. was detected in skin samples from two dogs, compatible with Leishmania (Viannia) braziliensis.
CONCLUSIONS/SIGNIFICANCE: Seroprevalence confirmed rapid increase in CVL in Puerto Iguazú. The rK28 RDT test potentially has great value for improved point-of-care diagnosis. Given cost reduction and accessibility, commercial LAMP may be applicable to buffy coat. RDT biomarkers of CVL clinical status are required to combat spread of CVL and HVL. The presence of Viannia, perhaps as an agent of human mucocutaneous leishmaniasis (MCL), highlights the need for vigilance and surveillance.
内脏利什曼病(VL)是一种由原生动物引起的人畜共患的虫媒传染病,是一项重大的公共卫生挑战。在阿根廷,犬内脏利什曼病(CVL)和人类内脏利什曼病(HVL)最近已经出现。目前缺乏针对 CVL 的标准化诊断检测方法,这阻碍了 CVL 和 HVL 的控制。
方法/主要发现:在阿根廷伊瓜苏港进行了采样,包括 190 只无症状、寡症状和多症状的狗。应用了以下诊断方法:淋巴结抽吸物(LNA)的显微镜检查;三种免疫层析快速诊断检测(RDT),原型 rK28-ICT、rK39-ICT(均来自 Coris BioConcept)、商业 rK39(InBios);针对 rK28、rK39 或粗裂解物抗原的 IgG、IgG1 和 IgG2 的 ELISA。使用皮肤样本对 30 只狗进行了 DNA 检测和分析,检测了 ITS1 区域,并用环介导等温扩增(Eiken Loopamp)对缓冲液、皮肤刮片或 LNA 进行检测。LNA 显微镜检查的阳性率为 15.4%。rK28 RDT 的血清阳性率(61%)高于原型 rK39 RDT(31.4%)或商业 rK39 RDT(18.8%),与其他六种病原体无交叉反应。IgG 抗 rK39 ELISA 抗体滴度,但不是 IgG2,与临床症状数量呈正相关。用 LNA 进行 LAMP 的阳性率高于 PCR;缓冲液取样比皮肤刮片更敏感。ITS1 证实 CVL 的病原体为利什曼原虫(Leishmania)婴儿。在两只狗的皮肤样本中检测到利什曼原虫(Viannia) spp.,与利什曼原虫(Viannia)巴西利什曼原虫相符。
结论/意义:血清阳性率证实了伊瓜苏港 CVL 的快速增长。rK28 RDT 检测法具有提高即时护理诊断的巨大价值。鉴于成本降低和可及性,商业 LAMP 可能适用于缓冲液。需要 CVL 临床状态的 RDT 生物标志物来对抗 CVL 和 HVL 的传播。Viannia 的存在,可能是人类黏膜皮肤利什曼病(MCL)的病原体,突显了保持警惕和监测的必要性。