FilmArray, an automated nested multiplex PCR system for multi-pathogen detection: development and application to respiratory tract infection.
FilmArray, an automated nested multiplex PCR system for multi-pathogen detection: development and application to respiratory tract infection.
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DOI：
10.1371/journal.pone.0026047
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Ririe KM
中科院分区：
文献类型：
--
作者：
Poritz MA;Blaschke AJ;Byington CL;Allen L;Nilsson K;Jones DE;Thatcher SA;Robbins T;Lingenfelter B;Amiott E;Herbener A;Daly J;Dobrowolski SF;Teng DH;Ririe KM
理想的临床诊断系统应提供快速、灵敏、特异和可重复的结果，同时最大限度地减少对专业实验室设施和熟练技术人员的要求。我们描述了一个集成的诊断平台，“FilmArray”，它完全自动化的检测和识别多种微生物从一个单一的样本在大约一个小时。对未处理的生物/临床样本进行核酸纯化、逆转录、高阶嵌套多重聚合酶链反应和扩增子熔解曲线分析。生化反应被封装在一次性袋中，最大限度地降低了PCR污染风险。FilmArray有可能一次检测超过100种不同的核酸靶标。这些特征使得该系统非常适合于感染因子的分子检测。通过开发一组能够鉴别21种常见病毒和细菌呼吸道病原体的检测试剂盒，实现了对FilmArray技术的验证。使用培养的微生物和从儿童获得的临床鼻抽吸物对该系统进行的初步测试证明了与现有诊断平台相当的分析和临床灵敏度和特异性。我们证明，可以通过分析扩增子的DNA解链曲线来实现从其相应的靶扩增子中自动鉴定病原体。
The ideal clinical diagnostic system should deliver rapid, sensitive, specific and reproducible results while minimizing the requirements for specialized laboratory facilities and skilled technicians. We describe an integrated diagnostic platform, the “FilmArray”, which fully automates the detection and identification of multiple organisms from a single sample in about one hour. An unprocessed biologic/clinical sample is subjected to nucleic acid purification, reverse transcription, a high-order nested multiplex polymerase chain reaction and amplicon melt curve analysis. Biochemical reactions are enclosed in a disposable pouch, minimizing the PCR contamination risk. FilmArray has the potential to detect greater than 100 different nucleic acid targets at one time. These features make the system well-suited for molecular detection of infectious agents. Validation of the FilmArray technology was achieved through development of a panel of assays capable of identifying 21 common viral and bacterial respiratory pathogens. Initial testing of the system using both cultured organisms and clinical nasal aspirates obtained from children demonstrated an analytical and clinical sensitivity and specificity comparable to existing diagnostic platforms. We demonstrate that automated identification of pathogens from their corresponding target amplicon(s) can be accomplished by analysis of the DNA melting curve of the amplicon.
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影响因子：
4.1
作者：
Dundas, Nicola E.;Ziadie, Mandolin S.;Rogers, Beverly B.
通讯作者：
Rogers, Beverly B.
影响因子：
9.4
作者：
Ginevra, C;Barranger, C;Grattard, F
通讯作者：
Grattard, F
影响因子：
9.4
作者：
Barenfanger, J;Drake, C;Troutt, T
通讯作者：
Troutt, T
DOI：
10.1016/s1386-6532(07)70006-9
发表时间：
2007-11
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Fox JD
通讯作者：
Fox JD
影响因子：
3.3
作者：
Charles, Patrick G. P.
通讯作者：
Charles, Patrick G. P.