'Real-time' PCR-based detection of Coxiella burnetii using conventional techniques

被引:13
作者
Frangoulidis, Dimitrios [1 ]
Meyer, Hermann [1 ]
Kahlhofer, Claudia [1 ]
Splettstoesser, Wolf D. [1 ]
机构
[1] Bundeswehr Inst Microbiol, D-80937 Munich, Germany
来源
FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY | 2012年 / 64卷 / 01期
关键词
Coxiella burnetii; PCR detection; real-time; improvement; deployable method;
D O I
10.1111/j.1574-695X.2011.00900.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The diagnosis of Q fever (Coxiella burnetii infection) relies primarily on the serological detection of specific antibodies. Recently, PCR-based methods have been introduced in diagnostic laboratories. Unfortunately, the fastest and most reliable real-time detection method, which employs the online detection of target nucleotide sequences while the amplification process is still in progress, requires expensive devices and consumables. In this study, we present a simple method that combines the simplicity of conventional PCR with new technical and methodical enhancements, resulting in a fast, specific and easy method for the molecular detection of C. burnetii. A collection of C. burnetii reference strains was tested with the modified conventional gel-based PCR approach applying a particluar PCR buffer (QIAGEN (R) Fast Cycling PCR kit) and using a closed ready-to-use gel-cassette-system (FlashGel (R)) for the visualization of specific PCR products. The modified conventional PCR method reached nearly the speed of the LightCycler (R) HybProbe real-time PCR assay (120 vs. 90 min) and showed equal sensitivity and specificity. The general cost per PCR run was 25% less than that for the LightCycler method. These improvements make this method suitable for small laboratories with limited resources and for deployable PCR diagnostics in field laboratories.
引用
收藏
页码:134 / 136
页数:3
相关论文
共 3 条
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[2]  
Scola Bernard La, 2002, Semin Pediatr Infect Dis, V13, P257
[3]  
Stemmler M, 2002, METHODS APPL MICROBI, P149