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Comparison of acid- versus heat-treatment for immune complex dissociation and detection of Dirofilaria immitis antigen in canine plasma.
Veterinary Parasitology ( IF 2.0 ) Pub Date : 2020-05-20 , DOI: 10.1016/j.vetpar.2020.109134
L A Starkey 1 , J V Bowles 1 , B L Blagburn 1
Affiliation  

Annual antigen testing is a mainstay for diagnosing infection with Dirofiliaria immitis in dogs; yet, it has been documented that some heartworm-infected dogs and cats test false-negative for antigen due to the presence of antigen-antibody complexes. Several studies have reported the use of heat as a reliable means of immune-complex dissociation (ICD) in recent years; however, the data regarding the use of acid as a reliable method of ICD for D. immitis detection are limited. The objective of this study was to compare an acid-based form of ICD to the more established and evaluated method of heat-based ICD in experimentally infected and non-infected dogs. Plasma from class A dogs experimentally infected ∼4 months prior with D. immitis (infected; n = 24) and dogs reared indoors with no history of exposure to mosquitoes (non-infected; n = 75) were evaluated for presence of D. immitis antigen (DiroCHEK® Heartworm antigen test kit). Each sample was divided into three aliquots for testing: [1] Control plasma (no acid- or heat-treatment), [2] acid-treated plasma (trichloroacetic acid (TCA), incubation, centrifugation for 5 min at 16,000 X g, buffer), and [3] heat-treated plasma (104 °C followed by centrifugation at 16,000 X g). Treatments for each aliquot were performed and tested in triplicate; results were determined both visually (color change) and by spectrophotometric analysis (optical density [OD] value). Of the 24 infected dogs, 0/24 tested positive for antigen in the absence of acid- or heat-treatment. Those same plasma samples following processing by either acid- or heat-treatment yielded 18/24 (75.0%) and 19/24 (79.2%) antigen-positive results, respectively. Of the 75 plasma samples from non-infected dogs, neither acid- nor heat-treatment of plasma caused any false-positive color changes or spectrophotometric values. These results indicate that acid as a means of ICD reliably allowed for the detection of D. immitis antigen in infected plasma while not inducing false-positive results in non-infected plasma samples.



中文翻译:

酸与热处理对免疫复合物解离的比较以及犬血浆中丝虫免疫力的检测。

年度抗原检测是诊断狗中Dirofiliaria炎的主要手段。然而,据报道,由于存在抗原-抗体复合物,一些感染了丝虫的狗和猫的抗原测试为假阴性。近年来有几项研究报告了使用热量作为免疫复合物解离(ICD)的可靠方法。然而,关于将酸用作ICD检测D.炎性炎的可靠方法的数据是有限的。这项研究的目的是在实验感染和未感染的犬只中,将一种以酸为基础的ICD与以热为基础的ICD的方法进行比较。从A级犬等离子实验感染与前几个月约为4 D.丝虫(感染; ñ = 24)和没有室内暴露于蚊子史(未感染;n  = 75)的室内饲养的狗被评估是否存在D. Immitis抗原(DiroCHEK®Heartworm抗原测试试剂盒)。每个样品分为三个等分试样进行测试:[1]对照血浆(不进行酸或热处理),[2]酸处理的血浆(三氯乙酸(TCA),孵育,以16,000 X g离心5分钟,缓冲液)和[3]热处理血浆(104°C,然后以16,000 X g离心))。每个等份的处理均进行三次,并进行测试;目测(颜色变化)和分光光度分析(光密度[OD]值)都确定了结果。在24只受感染的狗中,没有进行酸或热处理时0/24的抗原呈阳性。通过酸或热处理处理后的那些相同血浆样品分别产生18/24(75.0%)和19/24(79.2%)抗原阳性结果。在来自未感染狗的75个血浆样品中,血浆的酸化或热处理均未引起任何假阳性的颜色变化或分光光度值。这些结果表明,作为ICD手段的酸可靠地用于检测被感染血浆中的D.炎性炎症抗原,而不会在未感染血浆样品中引起假阳性结果。

更新日期:2020-05-20
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