A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection.
A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection.
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DOI：
10.3390/ijms17081250
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发表时间：
2016-08-03
影响因子：
5.6
通讯作者：
Hao Q
中科院分区：
文献类型：
--
作者：
Liu W;Liu HX;Zhang L;Hou XX;Wan KL;Hao Q
本研究建立了一种检测伯氏疏螺旋体（B. burgdorferi）的横向流动重组酶扩增等温检测方法（LF-RPA）。本试验灵敏度高，特异性强，无需任何额外设备，30分钟即可得到明显结果。根据伯氏疏螺旋体菌株的recA基因设计了一对引物，并进行了方法学评价。结果表明，基于recA基因的RPA检测方法成功应用于伯氏疏螺旋体检测，且只能从伯氏疏螺旋体基因组DNA中实现特异性扩增。新方法的检出限约为25个伯氏疏螺旋体基因组DNA拷贝。采用LF-RPA法、实时荧光定量pcr和巢式pcr对20例莱姆病患者血清进行检测。结果表明，LF-RPA法比巢式pcr法反应时间短，检出率高。该方法对莱姆病的临床快速检测具有重要价值。在临床和流行地区，使用RPA分析可能是DNA检测的大趋势。
A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients’ serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.
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影响因子：
7.4
作者：
Crannell, Zachary Austin;Castellanos-Gonzalez, Alejandro;Irani, Ayesha;Rohrman, Brittany;White, Arthur Clinton;Richards-Kortum, Rebecca
通讯作者：
Richards-Kortum, Rebecca
DOI：
10.1099/00207713-44-4-743
发表时间：
1994-10-01
期刊：
INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY
影响因子：
--
作者：
POSTIC, D;ASSOUS, MV;BARANTON, G
通讯作者：
BARANTON, G
影响因子：
4.4
作者：
Marques, Adriana R.
通讯作者：
Marques, Adriana R.
影响因子：
3.8
作者：
Chao CC;Belinskaya T;Zhang Z;Ching WM
通讯作者：
Ching WM
影响因子：
168.9
作者：
Stanek, Gerold;Wormser, Gary P.;Strle, Franc
通讯作者：
Strle, Franc