Comparative Sensitivity and Specificity of the 7SL sRNA Diagnostic Test for Animal Trypanosomiasis.
Comparative Sensitivity and Specificity of the 7SL sRNA Diagnostic Test for Animal Trypanosomiasis.
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DOI：
10.3389/fvets.2022.868912
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发表时间：
2022
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
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动物锥虫病（AT）是一种重要的牲畜疾病，影响撒哈拉以南非洲、中美洲和南美洲以及亚洲的数百万头牲畜，由原生动物寄生虫布鲁氏锥虫、间日锥虫和刚果锥虫引起，对牛的经济影响最大。由于现有诊断测试不充分，过度依赖推定化疗，这突出了改进AT诊断的必要性。一种小的RNA物种，7SL sRNA，由感染动物的锥虫排泄/分泌，并且先前已被证明可可靠地诊断活动性感染。我们试图探索7SL sRNA RT-qPCR检测的关键特性；即评估与广泛存在的良性锥虫交叉反应的可能性，直接将检测性能与现有诊断方法进行比较，定量评估特异性和敏感性，并评估7SL sRNA处理后的衰减率。结果表明，布鲁氏体、间日疟原虫和刚果螺旋体的7SL sRNA RT-qPCR检测效果优于显微镜和DNA PCR检测。7SL sRNA信号在处理96 h后无法检测到或显著降低；在治疗剂量为1倍的情况下，5/5感染刚果热锥虫的牛和3/5感染间日疟的牛没有检测到信号，其余2头感染间日疟的牛的信号减少了14630倍。此外，该测定法与T. theileri没有交叉反应。最后，通过大量验证的感染和未感染样本，通过受试者工作特征（ROC）分析显示，种特异性分析具有高度敏感性和特异性，对布鲁氏T具有100%的敏感性（95% CI, 96.44-100%）和100%的特异性（95% CI, 96.53-100%）， 96.73% （95% CI, 95.54-99.96%）和99.19%的特异性（95% CI, 92.58-99.60%）， 93.42% （95% CI, 85.51-97.16%）和82.43%的特异性（95% CI, 72.23-89.44%）。在使用的条件下，分别进行刚果弓形虫和间日弓形虫的测定。这些发现表明，7SL sRNA具有作为潜在的AT诊断标记物所需的许多特性：与氏肠杆菌无交叉反应、高特异性和敏感性、早期感染检测、即使血液中没有可检测到的寄生虫血症也能持续发出信号，以及在感染动物和治疗动物之间有明确的区分。
Animal trypanosomiasis (AT) is a significant livestock disease, affecting millions of animals across Sub-Saharan Africa, Central and South America, and Asia, and is caused by the protozoan parasites Trypanosoma brucei, Trypanosoma vivax, and Trypanosoma congolense, with the largest economic impact in cattle. There is over-reliance on presumptive chemotherapy due to inadequate existing diagnostic tests, highlighting the need for improved AT diagnostics. A small RNA species, the 7SL sRNA, is excreted/secreted by trypanosomes in infected animals, and has been previously shown to reliably diagnose active infection. We sought to explore key properties of 7SL sRNA RT-qPCR assays; namely, assessing the potential for cross-reaction with the widespread and benign Trypanosoma theileri, directly comparing assay performance against currently available diagnostic methods, quantitatively assessing specificity and sensitivity, and assessing the rate of decay of 7SL sRNA post-treatment. Results showed that the 7SL sRNA RT-qPCR assays specific for T. brucei, T. vivax, and T. congolense performed better than microscopy and DNA PCR in detecting infection. The 7SL sRNA signal was undetectable or significantly reduced by 96-h post treatment; at 1 × curative dose there was no detectable signal in 5/5 cattle infected with T. congolense, and in 3/5 cattle infected with T. vivax, with the signal being reduced 14,630-fold in the remaining two T. vivax cattle. Additionally, the assays did not cross-react with T. theileri. Finally, by using a large panel of validated infected and uninfected samples, the species-specific assays are shown to be highly sensitive and specific by receiver operating characteristic (ROC) analysis, with 100% sensitivity (95% CI, 96.44–100%) and 100% specificity (95% CI, 96.53–100%), 96.73% (95% CI, 95.54–99.96%) and 99.19% specificity (95% CI, 92.58–99.60%), and 93.42% (95% CI, 85.51–97.16% %) and 82.43% specificity (95% CI, 72.23–89.44% %) for the T brucei, T. congolense and T. vivax assays, respectively, under the conditions used. These findings indicate that the 7SL sRNA has many attributes that would be required for a potential diagnostic marker of AT: no cross-reaction with T. theileri, high specificity and sensitivity, early infection detection, continued signal even in the absence of detectable parasitaemia in blood, and clear discrimination between infected and treated animals.
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影响因子：
3.2
作者：
Cox AP;Tosas O;Tilley A;Picozzi K;Coleman P;Hide G;Welburn SC
通讯作者：
Welburn SC
影响因子：
2.6
作者：
Boulange, Alain;Pillay, Davita;Baltz, Theo
通讯作者：
Baltz, Theo
DOI：
10.20506/rst.34.2.2382
发表时间：
2015-08-01
影响因子：
0.6
作者：
Auty, H.;Torr, S. J.;Morrison, L. J.
通讯作者：
Morrison, L. J.
影响因子：
5.7
作者：
Brown, LD;Cai, TT;DasGupta, A
通讯作者：
DasGupta, A
影响因子：
3.2
作者：
Habeeb IF;Chechet GD;Kwaga JKP
通讯作者：
Kwaga JKP