Zysk G, Splettstösser W D, Neubauer H
Institute of Microbiology, Federal Armed Forces Medical Academy, Munich, Germany.
Clin Lab. 2000;46(3-4):119-30.
Melioidosis is an 'emerging' tropical disease which causes diagnostic problems in endemic and especially in nonendemic areas e.g. Germany when imported. In the last decade, many efforts have been made to develop new molecular and immunological techniques for diagnostic use. However, an actual comprehensive review does not exist. Apart from classical microbiological procedures (microscopy, culture and biochemical identification) efforts have been made to identify Burkholderia pseudomallei using specific antibodies and PCR. The direct antigen detection can be done within a few hours leading to diagnosis days before cultural proof. ELISA and immunoblot techniques were examined for their ability to replace indirect hemagglutination and immunofluorescence test in serology. The diagnostic value of serological procedures for early detection of melioidosis is limited, however. The rapid and reliable diagnosis of melioidosis is required for an adequate onset of therapy. But no evaluated test kit based on the detection of specific antibodies, specific antigens, or on the amplification of species-specific DNA sequences is commercially available up to now. Even PCR testing--primers can easily be ordered by many gene technology companies--can not be recommended as B. pseudomallei DNA for positive controls is not available. Therefore, only microscopy and biochemical identification systems like the API 20NE can be used in the routine laboratory up to now. At this point, it has to be stressed that B. pseudomallei is a level 3 agent in many countries e.g. Germany and that only laboratories with high containment are allowed to handle it. In all cases the help of an experienced reference laboratory is adviced.
类鼻疽是一种“新出现的”热带疾病,在流行地区尤其是非流行地区(如德国,当病例为输入性时)会引发诊断难题。在过去十年里,人们为开发用于诊断的新分子技术和免疫技术付出了诸多努力。然而,目前尚无实际的全面综述。除了经典的微生物学程序(显微镜检查、培养和生化鉴定)外,人们还尝试使用特异性抗体和聚合酶链反应(PCR)来鉴定类鼻疽杆菌。直接抗原检测可在数小时内完成,比培养确诊提前数天得出诊断结果。酶联免疫吸附测定(ELISA)和免疫印迹技术已被检验其在血清学中替代间接血凝试验和免疫荧光试验的能力。不过,血清学检测程序对类鼻疽早期检测的诊断价值有限。为了能适时开始治疗,需要对类鼻疽进行快速可靠的诊断。但截至目前,尚无基于检测特异性抗体、特异性抗原或扩增物种特异性DNA序列的经评估的诊断试剂盒可供商业使用。即便PCR检测——许多基因技术公司都能轻易订购引物——也不被推荐,因为无法获得用于阳性对照的类鼻疽杆菌DNA。因此,到目前为止,常规实验室只能使用显微镜检查和生化鉴定系统,如API 20NE。在此必须强调,在许多国家(如德国),类鼻疽杆菌属于3级病原体,只有具备高防护水平的实验室才允许处理它。在所有情况下,都建议寻求经验丰富的参考实验室的帮助。