Abstract：
Pythiosis, a life-threatening infectious condition caused by Pythium insidiosum, has been increasingly reported in humans and animals worldwide. Antifungal drugs usually fail to control the pathogen. The surgical removal of an infected organ is the treatment of choice. Many affected patients die due to advanced infection. A timely and accurate diagnosis could lead to a better prognosis in pythiosis patients and save their lives. Although a standard culture method is available in microbiological laboratories, it is time-consuming, laborious, and insensitive for P. insidiosum identification. Immunological assays have been developed to improve the diagnosis of pythiosis. However, immunological methods are commercially unavailable and primarily detect anti-P. insidiosum antibodies, which constitute indirect evidence of pythiosis, making it challenging to differentiate a past from a recent infection. Moreover, such immunological tests cannot diagnose patients with a local infection, such as in the eye. Nucleic acid-based tests (NATs) are efficient for the direct and rapid detection of P. insidiosum DNA in trace-amount or culture-negative specimens. The reagents and equipment required for NATs are usually available in molecular diagnostic laboratories. Herein, we provide a systematic review to comprehensively present the principal and clinical usages, advantages, and limitations of such NATs in the detection of P. insidiosum. Various NATs have been established to detect P. insidiosum, which can be classified into amplification-based (i.e., PCR assays, isothermal tests, and next-generation sequencing methods) and non-amplification-based (i.e., DNA hybridization) techniques. This concise review on NATs constitutes an up-to-date reference with which healthcare professionals can learn about and decide upon which detection method is suitable for their respective laboratory environments.
摘要：
脓疱,一种由泥腐菌引起的危及生命的传染病，在世界各地的人类和动物中越来越多的报道。抗真菌药物通常无法控制病原体。手术切除受感染的器官是选择的治疗方法。许多受影响的患者死于晚期感染。及时准确的诊断可使患者预后较好,挽救生命。尽管微生物实验室有标准的培养方法，这是耗时的，辛苦,对山药鉴定不敏感。已经开发了免疫测定法以改善化脓症的诊断。然而,免疫学方法在商业上不可用，主要检测抗P。阴草抗体，构成化脓症的间接证据，很难区分过去和最近的感染。此外，这种免疫检测不能诊断局部感染的患者，比如眼睛。基于核酸的测试（NAT）可有效地直接快速检测痕量或培养阴性标本中的阴阳假单胞菌DNA。NAT所需的试剂和设备通常可在分子诊断实验室获得。在这里，我们提供了一个系统的审查，以全面介绍的主要和临床用法，优势,以及此类NAT在检测阴阳假单胞菌中的局限性。已经建立了各种NAT来检测阴间假单胞菌，可以分为基于放大的(即，PCR检测,等温试验,和下一代测序方法)和非基于扩增的(即，DNA杂交)技术。对NAT的简要审查构成了最新的参考，医疗保健专业人员可以通过该参考来了解并决定哪种检测方法适合其各自的实验室环境。
