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感染肺线虫隐匿猫圆线虫的猫体内特异性抗体的检测。

Detection of specific antibodies in cats infected with the lung nematode Aelurostrongylus abstrusus.

作者信息

Zottler Eva-Maria, Strube Christina, Schnyder Manuela

机构信息

University of Zurich, Vetsuisse Faculty, Institute of Parasitology, Winterthurerstrasse 266a, 8057 Zurich, Switzerland.

University of Veterinary Medicine Hannover, Institute for Parasitology, Centre for Infection Medicine, Buenteweg 17, 30559 Hannover, Germany.

出版信息

Vet Parasitol. 2017 Feb 15;235:75-82. doi: 10.1016/j.vetpar.2017.01.015. Epub 2017 Jan 17.

Abstract

Feline aelurostrongylosis, caused by the metastrongylid nematode Aelurostrongylus abstrusus, is an underestimated respiratory parasitosis. Its diagnosis currently mainly relies on the isolation of first stage larvae from fresh faecal samples. The aim of our study was to develop a serological test for the detection of specific antibodies against A. abstrusus by ELISA. We used recombinant major sperm protein (MSP) of the bovine lungworm Dictyocaulus viviparus as detection antigen and evaluated two different ELISA plates (Maxisorp and Immobilizer™ Amino-plate, Nunc Roskilde, Denmark) with two different enzyme systems [alkaline phosphatase (AP) and horseradish peroxidase (HRP)]. Sera from cats experimentally (n=54) and naturally (n=17) infected with A. abstrusus and from randomly selected cats with different medical issues (n=160) were used to determine sensitivity and specificity. Furthermore, cross-reactions were evaluated using sera from cats naturally (n=71) and experimentally (n=8) infected with different nematodes. A sensitivity of 100% was obtained with sera from experimentally infected cats at 10 weeks post infection using MSP on the Immobilizer™ Amino-plate with HRP, while it ranged between 90.5 and 95.2% in the other ELISA set-ups. Using sera from naturally infected cats, a sensitivity of 88.2% (95% confidence interval: 63.6-98.5%) was achieved in all four set-ups. The specificity was 85.2-94.4% in sera from uninfected cats prior to experimental infection and 68.1-90% in randomly selected cats depending on the plate and enzyme system. The number of seropositive cats increased over time post infection. Serological follow-up showed a decrease of antibody levels within 30days after anthelmintic treatment. Seropositive reactions were observed with sera from stray cats naturally infected with Toxocara cati, Capillaria sp., hookworms and Taeniidae; however, coproscopic false negative A. abstrusus findings cannot be excluded. The serological detection of specific antibodies against A. abstrusus using ELISA requires a single serum sample and therefore represents a valid alternative for reliable individual diagnosis of A. abstrusus in cats and facilitates mass screening, overcoming the usually difficult collection of cat faeces.

摘要

猫肺并殖吸虫病由后圆线虫纲线虫——隐匿猫肺并殖吸虫引起,是一种未得到充分重视的呼吸道寄生虫病。目前其诊断主要依靠从新鲜粪便样本中分离出第一期幼虫。我们研究的目的是开发一种酶联免疫吸附测定(ELISA)法,用于检测针对隐匿猫肺并殖吸虫的特异性抗体。我们使用牛肺线虫——胎生网尾线虫的重组主要精子蛋白(MSP)作为检测抗原,并评估了两种不同的ELISA板(Maxisorp板和Immobilizer™氨基板,丹麦罗斯基勒的Nunc公司)以及两种不同的酶系统[碱性磷酸酶(AP)和辣根过氧化物酶(HRP)]。使用来自实验感染(n = 54)和自然感染(n = 17)隐匿猫肺并殖吸虫的猫的血清,以及随机选择的患有不同疾病的猫(n = 160)的血清来确定敏感性和特异性。此外,使用来自自然感染(n = 71)和实验感染(n = 8)不同线虫的猫的血清评估交叉反应。使用HRP标记的Immobilizer™氨基板上的MSP,在感染后10周时,实验感染猫的血清获得了100%的敏感性,而在其他ELISA设置中,敏感性在90.5%至95.2%之间。对于自然感染猫的血清,在所有四种设置中敏感性达到了88.2%(95%置信区间:63.6 - 98.5%)。在实验感染前未感染猫的血清中,特异性为85.2 - 94.4%,在随机选择的猫中,根据板和酶系统的不同,特异性为68.1 - 90%。感染后血清学阳性猫的数量随时间增加。血清学随访显示驱虫治疗后30天内抗体水平下降。在自然感染猫弓首蛔虫、毛细属线虫、钩虫和带绦虫的流浪猫的血清中观察到血清学阳性反应;然而,不能排除粪便检查中隐匿猫肺并殖吸虫假阴性结果。使用ELISA法对隐匿猫肺并殖吸虫特异性抗体进行血清学检测只需要一份血清样本,因此是对猫隐匿猫肺并殖吸虫进行可靠个体诊断的有效替代方法,有助于大规模筛查,克服了通常难以收集猫粪便的问题。

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