Department of Veterinary Medicine, University of Nigeria, Nsukka, Nigeria.
Department of Biochemistry, Genetics and Microbiology,University of Pretoria, Pretoria, South Africa.
PLoS Negl Trop Dis. 2020 Feb 28;14(2):e0008088. doi: 10.1371/journal.pntd.0008088. eCollection 2020 Feb.
Rabies lyssavirus (RABV) is the aetiologic agent of rabies, a disease that is severely underreported in Nigeria as well as elsewhere in Africa and Asia. Despite the role that rabies diagnosis plays towards elucidating the true burden of the disease, Nigeria-a country of 180 million inhabitants-has a limited number of diagnostic facilities. In this study, we sought to investigate two of the World Organization for Animal Health (OIE)-recommended diagnostic assays for rabies-viz; the direct fluorescent antibody test (DFA) and the direct rapid immunohistochemical test (dRIT) in terms of their relative suitability in resource-limited settings. Our primary considerations were (1) the financial feasibility for implementation and (2) the diagnostic efficacy. As a case study, we used suspect rabies samples from dog meat markets in Nigeria.
METHODS/PRINCIPAL FINDINGS: By developing a simple simulation framework, we suggested that the assay with the lowest cost to implement and routinely use was the dRIT assay. The costs associated with the dRIT were lower in all simulated scenarios, irrespective of the number of samples tested per year. In addition to the cost analysis, the diagnostic efficacies of the two assays were evaluated. To do this, a cohort of DFA-positive and -negative samples collected from dog meat markets in Nigeria were initially diagnosed using the DFA in Nigeria and subsequently sent to South Africa for diagnostic confirmation. In South Africa, all the specimens were re-tested with the DFA, the dRIT and a quantitative real-time polymerase chain reaction (qRT-PCR). In our investigation, discrepancies were observed between the three diagnostic assays; with the incongruent results being resolved by means of confirmatory testing using the heminested reverse transcription polymerase reaction and sequencing to confirm that they were not contamination.
CONCLUSIONS/SIGNIFICANCE: The data obtained from this study suggested that the dRIT was not only an effective diagnostic assay that could be used to routinely diagnose rabies, but that the assay was also the most cost-effective option among all of the OIE recommended methods. In addition, the results of our investigation confirmed that some of the dogs slaughtered in dog markets were rabies-positive and that the markets posed a potential public health threat. Lastly, our data showed that the DFA, although regarded as the gold standard test for rabies, has some limitations-particularly at low antigen levels. Based on the results reported here and the current challenges faced in Nigeria, we believe that the dRIT assay would be the most suitable laboratory test for decentralized or confirmatory rabies diagnosis in Nigeria, given its relative speed, accuracy, cost and ease of use.
狂犬病病毒(Rabies lyssavirus,RABV)是狂犬病的病原体,这种疾病在尼日利亚以及非洲和亚洲的其他地方严重漏报。尽管狂犬病诊断在阐明疾病的真实负担方面发挥了作用,但尼日利亚这个拥有 1.8 亿人口的国家,其诊断设施数量有限。在这项研究中,我们试图研究世界动物卫生组织(World Organization for Animal Health,OIE)推荐的两种狂犬病诊断检测方法,即直接荧光抗体检测(direct fluorescent antibody test,DFA)和直接快速免疫组织化学检测(direct rapid immunohistochemical test,dRIT),以评估它们在资源有限的环境中的相对适用性。我们的主要考虑因素是(1)实施的财务可行性,以及(2)诊断效果。作为一个案例研究,我们使用了来自尼日利亚狗肉市场的可疑狂犬病样本。
方法/主要发现:通过开发一个简单的模拟框架,我们建议实施成本最低且常规使用的检测方法是 dRIT 检测。在所有模拟场景中,dRIT 的相关成本都较低,而与每年测试的样本数量无关。除了成本分析外,我们还评估了这两种检测方法的诊断效果。为此,我们首先使用来自尼日利亚狗肉市场的 DFA 阳性和阴性样本进行了队列研究,然后将这些样本送到南非进行诊断确认。在南非,所有样本均使用 DFA、dRIT 和实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)进行了重新检测。在我们的研究中,三种诊断检测方法之间存在差异;通过使用半巢式逆转录聚合酶链反应(heminested reverse transcription polymerase reaction)和测序进行确认性检测,解决了不一致的结果,以确认它们不是污染。
结论/意义:这项研究的结果表明,dRIT 不仅是一种有效的诊断方法,可以常规用于诊断狂犬病,而且是所有 OIE 推荐方法中最具成本效益的选择。此外,我们的调查结果证实,一些在狗肉市场屠宰的狗患有狂犬病,这些市场对公共健康构成了潜在威胁。最后,我们的数据表明,尽管 DFA 被认为是狂犬病的金标准检测方法,但它存在一些局限性,尤其是在抗原水平较低的情况下。基于这里报告的结果以及尼日利亚目前面临的挑战,我们认为,鉴于其相对速度、准确性、成本和易用性,dRIT 检测方法将是尼日利亚进行去中心化或确认性狂犬病诊断的最合适的实验室检测方法。