Cancrini G, Iori A
Dip. di Scienze di Sanità Pubblica, Sez. Parassitologia, Servizio di Analisi parassitologiche Policlinico Umberto I, Università La Sapienza, Roma.
Parassitologia. 2004 Jun;46(1-2):173-6.
The development of new technological methods surely improves the quality of the Diagnostic Services in Parasitology offered to the National Sanitary Service, however, cost and simplicity have not to be neglected, even when the prime consideration is efficiency. Moreover, the mere fact that something can be done by one of these new approaches does not mean that it should be done that way or that it is most cost-effective to do it that way. A review of diagnostic tools in Parasitology is proposed, to evaluate when and why each of them should be applied. Traditional procedures for the diagnosis of parasitosis are only based on the "direct" recovery and recognition of the parasite, with the microscope as main tool and few other instruments as co-operator. The innovative procedures, recently adjusted on the basis of new scientific knowledge and made possible by the development of the laboratory instrument weapons, can evidence the parasites both directly and indirectly. If it is obvious that the direct identification of a pathogen is more reliable than that indirect, is not so evident what is the most useful direct method, and when it would be better to use indirect diagnostic tools. Advantages and disadvantages of each procedure, cost as well as the purpose of the test (diagnosis, post-treatment, research), and the general condition in which the test have to been applied must be taken into account when we are choosing. In general, we can say that the rationale for their use can be summarised as follows: 1) The macroscopic/microscopic analysis of samples is always recommended (with the exception of samples coming from tissues that need surgery). This "old" procedure allows the identification in 20 minutes of all the parasites present in mixed infections, and the evaluation of the parasite load. It is a cost-effective method which relies ultimately on the skill of the observer to detect and identify parasite stages; 2) Parasite antigen detection is an innovative and expensive immunological diagnostic, which can suffer of sensitivity and specificity. It could be useful to directly diagnose "occult" infections; 3) Parasite DNA/RNA direct detection is an innovative, sensitive and specific procedure, which can also identify sibling species. It is expensive, therefore its use is restricted to reference laboratories; 4) Host antibody detection is an innovative indirect tool to evaluate the presence of a parasite by means the evaluation of the host response to infection. It can suffer of sensitivity and specificity, and the interpretation of the test results may be difficult. It could be applied as first step to evaluate the presence of tissue parasites, whose direct diagnosis would require surgery. Some tests can be performed in well-equipped laboratories; other tests are available through research laboratories. The specimens, appropriately collected and preserved, have always to be processed in security for potential risk of infection hazard, and submitted to tests appropriate to the laboratory's goals, where, therefore, field and research diagnostic tools shouldn't be applied. The test selected for routine use has to be chosen taking into account value and limitations of each method. Reduction in excessive and often unnecessary testing is mandatory, and therefore it is critical for the clinical Parasitology to perform relevant testing while maintaining appropriate quality. To date, the microscopic analysis of samples is the only direct method that allows all identifications in short times, at a reduced cost, independently from geographical origin and peculiar status of the patient. It has to be regarded as the first step in diagnostic procedures for all laboratories. Some molecular techniques have greater sensitivity than traditional methods, but at least at the present time, their costs may well preclude their routine use. It is difficult to know, exactly, where diagnostic Parasitology will be moving in the next few years, although many soothsayers feel very strongly that the area of molecular diagnostics will replace more traditional means. It is also possible that immunological or perhaps cytometric procedures will replace our more standard diagnostic approach; nevertheless they will continue to remain oddities on the outside of the general practice and be confined to a few reference laboratories. As far as semi-automated or automated instruments and robotics-based techniques, they are useful when large numbers of the same test are performed. Supposing that they will enter in our laboratory, that will happen in central facility rather than in each local facility. So, the great interest in using new technological methods to solve old problems probably will have to be seen in the right perspective.
新技术方法的发展无疑提高了国家卫生服务机构所提供的寄生虫学诊断服务的质量,然而,即便首要考虑的是效率,成本和简便性也不容忽视。此外,能用这些新方法之一完成某件事,并不意味着就应该那样做,或者那样做就是最具成本效益的。本文对寄生虫学诊断工具进行综述,以评估何时以及为何应应用每种工具。寄生虫病的传统诊断程序仅基于寄生虫的“直接”发现和识别,主要工具是显微镜,其他仪器为辅。基于新科学知识最近调整并借助实验室仪器发展得以实现的创新程序,能够直接和间接证实寄生虫的存在。显然,直接鉴定病原体比间接鉴定更可靠,但哪种直接方法最有用,以及何时最好使用间接诊断工具,却并不那么明显。选择时必须考虑每种程序的优缺点、成本以及检测目的(诊断、治疗后、研究),以及进行检测的一般条件。总体而言,其使用的基本原理可总结如下:1)始终建议对样本进行宏观/微观分析(来自需要手术的组织的样本除外)。这种“古老”程序能在20分钟内鉴定混合感染中存在的所有寄生虫,并评估寄生虫负荷。这是一种具有成本效益的方法,最终依赖于观察者检测和识别寄生虫阶段的技能;2)寄生虫抗原检测是一种创新且昂贵的免疫诊断方法,可能存在敏感性和特异性问题。它可能有助于直接诊断“隐匿”感染;3)寄生虫DNA/RNA直接检测是一种创新、敏感且特异的程序,也能识别近缘种。它成本高昂,因此其使用仅限于参考实验室;4)宿主抗体检测是一种创新的间接工具,通过评估宿主对感染的反应来评估寄生虫的存在。它可能存在敏感性和特异性问题,检测结果的解读可能很困难。它可作为评估组织寄生虫存在的第一步,其直接诊断需要手术。一些检测可在设备完善的实验室进行;其他检测可通过研究实验室获得。适当采集和保存的标本,由于存在感染风险,始终必须安全处理,并提交适合实验室目标的检测,因此,现场和研究诊断工具不应在那里使用。选择用于常规使用的检测时,必须考虑每种方法的价值和局限性。减少过度且往往不必要的检测是必要的,因此,临床寄生虫学在保持适当质量的同时进行相关检测至关重要。迄今为止,样本的显微镜分析是唯一能在短时间内、以较低成本、不受患者地理来源和特殊状况影响进行所有鉴定的直接方法。它应被视为所有实验室诊断程序的第一步。一些分子技术比传统方法具有更高的敏感性,但至少目前,其成本可能使其无法常规使用。很难确切知道未来几年诊断寄生虫学将走向何方,尽管许多预言家坚信分子诊断领域将取代更传统的方法。免疫或细胞计数程序也有可能取代我们更标准的诊断方法;然而,它们在一般实践之外仍将是异类,仅限于少数参考实验室。至于半自动或自动仪器以及基于机器人技术的方法,当进行大量相同检测时它们很有用。假设它们进入我们的实验室,那将发生在中心机构而非每个地方机构。所以,对使用新技术方法解决老问题的极大兴趣,或许需要从正确的角度来看待。