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不同单重和多重核酸扩增检测试剂在多病原体外部质量评估panel 上的表现。

Performance of different mono- and multiplex nucleic acid amplification tests on a multipathogen external quality assessment panel.

机构信息

Department of Microbiology, Vaccine and Infectious Diseases Institute, University of Antwerp, Antwerp, Belgium.

出版信息

J Clin Microbiol. 2012 Mar;50(3):977-87. doi: 10.1128/JCM.00200-11. Epub 2011 Dec 14.


DOI:10.1128/JCM.00200-11
PMID:22170925
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3295136/
Abstract

An external quality assessment (EQA) panel consisting of a total of 48 samples in bronchoalveolar lavage (BAL) fluid or transport medium was prepared in collaboration with Quality Control for Molecular Diagnostics (QCMD) (www.qcmd.org). The panel was used to assess the proficiency of the three laboratories that would be responsible for examining the 6,000 samples to be collected in the GRACE Network of Excellence (www.grace-lrti.org). The main objective was to decide on the best-performing testing approach for the detection of influenza viruses A and B, parainfluenza virus types 1 to 3, respiratory syncytial virus (RSV), human metapneumovirus, coronavirus, rhinovirus, adenovirus, Chlamydophila pneumoniae, Mycoplasma pneumoniae, and Legionella pneumophila by nucleic acid amplification techniques (NAATs). Two approaches were chosen: (i) laboratories testing samples using their in-house procedures for extraction and amplification and (ii) laboratories using their in-house amplification procedures on centrally extracted samples. Furthermore, three commercially available multiplex NAAT tests-the ResPlex (Qiagen GmbH, Hilden, Germany), RespiFinder plus (PathoFinder, Maastricht, The Netherlands), and RespiFinder Smart 21 (PathoFinder) tests-were evaluated by examination of the same EQA panel by the manufacturer. No large differences among the 3 laboratories were noticed when the performances of the assays developed in-house in combination with the in-house extraction procedures were compared. Also, the extraction procedure (central versus local) had little effect on performance. However, large differences in amplification efficacy were found between the commercially available tests; acceptable results were obtained by using the PathoFinder assays.

摘要

与分子诊断质量控制(Quality Control for Molecular Diagnostics,QCMD)(www.qcmd.org)合作,制备了总共 48 个支气管肺泡灌洗液(bronchoalveolar lavage,BAL)或运输介质的外部质量评估(EQA)试剂盒。该试剂盒用于评估将负责检查卓越研究网络(GRACE Network of Excellence,www.grace-lrti.org)中收集的 6000 个样本的三个实验室的熟练程度。主要目标是确定用于检测流感病毒 A 和 B、副流感病毒 1 至 3 型、呼吸道合胞病毒(respiratory syncytial virus,RSV)、人偏肺病毒、冠状病毒、鼻病毒、腺病毒、肺炎衣原体、肺炎支原体和嗜肺军团菌的最佳检测方法通过核酸扩增技术(nucleic acid amplification techniques,NAATs)。选择了两种方法:(i)使用内部提取和扩增程序测试样本的实验室,以及(ii)使用内部扩增程序对集中提取的样本进行测试的实验室。此外,还评估了三种市售的多重 NAAT 测试——ResPlex(Qiagen GmbH,德国希尔德斯海姆)、RespiFinder plus(PathoFinder,荷兰马斯特里赫特)和 RespiFinder Smart 21(PathoFinder)测试——通过制造商对相同的 EQA 试剂盒进行检测。当比较内部开发的检测方法与内部提取程序相结合的性能时,三个实验室之间没有发现明显差异。此外,提取程序(集中与本地)对性能的影响很小。然而,市售测试之间的扩增效果存在很大差异;使用 PathoFinder 检测可以获得可接受的结果。

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本文引用的文献

[1]
Evaluation of multiple commercial molecular and conventional diagnostic assays for the detection of respiratory viruses in children.

Clin Microbiol Infect. 2011-6-24

[2]
Evaluation of commercial ResPlex II v2.0, MultiCode-PLx, and xTAG respiratory viral panels for the diagnosis of respiratory viral infections in adults.

J Clin Virol. 2010-11-2

[3]
Molecular quantification of respiratory syncytial virus in respiratory samples: reliable detection during the initial phase of infection.

J Clin Microbiol. 2010-7-21

[4]
Acute respiratory infection due to Mycoplasma pneumoniae: current status of diagnostic methods.

Eur J Clin Microbiol Infect Dis. 2010-6-6

[5]
Disease severity and viral load are correlated in infants with primary respiratory syncytial virus infection in the community.

J Med Virol. 2010-7

[6]
A multicenter pilot external quality assessment programme to assess the quality of molecular detection of Chlamydophila pneumoniae and Mycoplasma pneumoniae.

J Microbiol Methods. 2010-5-19

[7]
Detection of respiratory viruses by molecular methods.

Clin Microbiol Rev. 2008-10

[8]
Evaluation of different nucleic acid amplification techniques for the detection of M. pneumoniae, C. pneumoniae and Legionella spp. in respiratory specimens from patients with community-acquired pneumonia.

J Microbiol Methods. 2008-6

[9]
Acute respiratory infection due to Chlamydia pneumoniae: current status of diagnostic methods.

Clin Infect Dis. 2007-2-15

[10]
Evaluation of NucliSens easyMAG for automated nucleic acid extraction from various clinical specimens.

J Clin Microbiol. 2007-2

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