Comparative assessment of conventional PCR with multiplex real-time PCR using SYBR Green I detection for the molecular diagnosis of imported malaria
Comparative assessment of conventional PCR with multiplex real-time PCR using SYBR Green I detection for the molecular diagnosis of imported malaria
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DOI：
10.1017/s0031182003004219
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发表时间：
2004-01-01
期刊：
影响因子：
2.4
通讯作者：
Magnaval, JF
中科院分区：
文献类型：
--
作者：
Fabre, R;Berry, A;Magnaval, JF
对于输入性疟疾的诊断，已知光学或免疫层析方法的敏感性和特异性低于基于pcr的方法，后者反过来更复杂和耗时。因此，采用了一种基于顺序使用多重竞争性实时PCR检测恶性疟原虫或疟原虫感染的原始策略，然后在必要时使用单一实时PCR进行物种鉴定，然后在常规条件下与常规PCR进行比较。自1999年10月以来，传统PCR已在图卢兹大学医院寄生虫科作为二线诊断方法使用。在183名接受检测的患者中，经常规显微镜检查发现48人感染恶性疟原虫，经常规PCR检测发现60人感染恶性疟原虫，经多重竞争性实时PCR检测发现60人感染恶性疟原虫。另外9例患者为非恶性疟原虫感染，常规PCR和单次实时PCR鉴定的种类一致。与显微镜技术相比，基于pcr的方法的主要价值在于确定可疑样品的阴性。此外，实时PCR简化了操作程序，可在2小时内做出诊断。
For the diagnosis of imported malaria, optical or immunochromatographic methods are known to be less sensitive and less specific than PCR-based methods, which are conversely more complicated and time-consuming. An original strategy, based upon the sequential use of a multiplex competitive real-time PCR detecting Plasmodium falciparum or Plasmodium spp. infection, followed by, if necessary, a single real-time PCR for species identification, was therefore performed and then tested versus conventional PCR in routine conditions. Conventional PCR has been used since October 1999 in the Department of Parasitology, University Hospitals in Toulouse, as a 2nd line diagnostic method. Out of 183 patients tested, 48 were found to be harbouring a falciparum infection by conventional microscopy, 60 by conventional PCR and 60 by multiplex competitive real-time PCR. Nine further patients had a non-falciparum infection, and concordant species identifications were obtained by both conventional PCR and single real-time PCR. The major value of PCR-based methods, when compared to microscopical techniques, was to ascertain the negativity of a suspect sample. Moreover, real-time PCR allows simplification of the operating procedure, with a diagnosis being made within 2 h.