Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp.
Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp.
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DOI：
10.4269/ajtmh.2007.76.713
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发表时间：
2007-04-01
影响因子：
3.3
通讯作者：
Petri, William A., Jr.
中科院分区：
文献类型：
--
作者：
Haque, Rashidul;Roy, Shantanu;Petri, William A., Jr.
溶组织内阿米巴、肠贾第虫和隐孢子虫属。不仅是三种最重要和最常见的引起腹泻的寄生原虫，而且它们通常具有相似的临床表现。这些寄生虫的显微镜诊断既不敏感也不特异。最近，针对所有这三种寄生虫感染引入了更特异和更灵敏的替代分子方法（聚合酶链反应 [PCR] 和抗原检测测试）。这些分子诊断测试在常规诊断实验室中的使用仍然有限。在本研究中，我们开发了一种多重实时 PCR 检测方法，用于同时检测溶组织内阿米巴、肠小球菌和隐孢子虫属。在使用物种特异性探针的一个反应中。使用临床标本对该测定法进行了评估，发现该测定法相当灵敏且特异。该多重 PCR 测定中使用的试剂也可用于单独检测这些寄生虫。目前，这种实时PCR多重检测在发展中国家的常规诊断范围有限，因为单次检测的成本很高，但在发达国家，该检测可以立即应用。
Entarnoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. are not only three of the most important and common diarrhea-causing parasitic protozoa, but they often have similar clinical presentations. Microscopic diagnosis of these parasites is neither sensitive nor specific. Recently, more specific and sensitive alternative molecular methods (polymerase chain reaction [PCR] and antigen detection tests) have been introduced for all three of these parasitic infections. The use of these molecular diagnostic tests in routine diagnostic laboratories is still limited. In this study, we developed a multiplex real-time PCR assay for the simultaneous detection of E. histolytica, G. intestinalis, and Cryptosporidium spp. in one reaction using species-specific probes. This assay was evaluated using clinical specimens and was found to be quite sensitive and specific. The reagents used in this multiplex PCR assay can also be used for detection of these parasites individually. The use of this real-time PCR multiplex assay in developing countries at present will have limited scope for routine diagnosis because the cost will be high for a single test, although in the developed world, the test could see immediate application.