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A Leishmania infantum FML-ELISA for the Detection of Symptomatic and Asymptomatic Canine Visceral Leishmaniasis in an Endemic Area of Iran

机译:婴儿利什曼原虫FML-ELISA用于检测伊朗流行区有症状和无症状的内脏利什曼病

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Background: Visceral leishmaniasis (VL) is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis (CVL). Objective: In this survey a Fucose-Mannose Ligand (FML) ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test (DAT) and PCR was evaluated. Methods: Blood samples of sixty ownership dogs (≤ 3 years old) were collected from Meshkin-shahr district in Ardabil province, North-west of Iran. Sera were separated for serological assays (DAT and FMLELISA) and the buffy coats were collected for molecular evaluation. Results: Two out of the 60 (3.33%) samples were found to be positive (antibody titer of ≥ 1/320) in DAT while seven of the 60 (11.66%) samples were positive by FML-ELISA. Nine out of 60 (15%) buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers 1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively. Conclusion: Considering the acceptable sensitivity and high specificity of FMLELISA, use of this serological method can be recommended for epidemiological surveys of CVL.
机译:背景:内脏利什曼病(VL)是由地中海盆地的婴儿利什曼原虫引起的,是伊朗某些地区的地方病。在大多数流行地区,犬科动物是VL的主要蓄水池。已经引入了不同的血清学方法来诊断犬内脏利什曼病(CVL)。目的:在这项调查中,使用天然婴儿乳杆菌抗原开发了岩藻糖-甘露糖配体(FML)ELISA,并与直接凝集试验(DAT)和PCR相比,评价了其对感染狗的检测有效性。方法:从伊朗西北部阿尔达比勒省的Meshkin-shahr地区收集了60只三岁以下的犬的血样。分离血清用于血清学测定(DAT和FMLELISA),并收集血沉棕黄层用于分子评估。结果:60份样品中有2份(3.33%)在DAT中呈阳性(抗体滴度≥1/320),而60份样品中有7份(11.66%)通过FML-ELISA呈阳性。在60个(15%)的血沉棕黄层样本中,有9个显示出一条约680 bp的条带,指示PCR中的婴儿乳杆菌。 60只狗中有3只具有Kala-azar症状,并且通过PCR和FML-ELISA呈阳性,而这三只狗中有两只在其血清样品中抗体滴度> 1/320。 FML-ELISA在有症状和无症状犬中检测CVL的敏感性和特异性分别为77.8%和100%。结论:考虑到FMLELISA的可接受的敏感性和高特异性,可以推荐使用这种血清学方法进行CVL的流行病学调查。

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