Development and laboratory evaluation of a real-time PCR assay for detecting viruses and bacteria of relevance for community-acquired pneumonia

The Journal of Molecular Diagnostics : JMD
Alicia EdinAnders Johansson

Abstract

Community-acquired pneumonia may present with similar clinical symptoms, regardless of viral or bacterial cause. Diagnostic assays are needed to rapidly discriminate between causes, because this will guide decisions on appropriate treatment. Therefore, a quantitative real-time PCR (qPCR) assay with duplex reactions targeting eight bacteria and six viruses was developed. Technical performance was examined with linear plasmids. Upper and lower respiratory tract specimens were used to compare the qPCR assay with standard microbiological methods. The limit of detection was 5 to 20 DNA template copies with approximately 1000-fold differences in concentrations of the two competing templates. SDs for positive controls were <5%. The use of the qPCR assay resulted in 113 positive identifications in 94 respiratory specimens compared with 38 by using standard diagnostics. Diagnostic accuracy of the qPCR assay varied between 60% positive agreement with standard tests for Streptococcus pneumoniae and 100% for Mycoplasma pneumoniae, Moraxella catarrhalis, and Staphylococcus aureus. Negative percentage of agreement was >95% for M. pneumoniae, Streptococcus pyogenes, respiratory syncytial virus, and influenza A virus; whereas it was only 56% f...Continue Reading

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Citations

Jan 10, 2016·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Naomi J GadsbyKate E Templeton
Apr 12, 2016·Frontiers in Microbiology·Katherine Loens, Margareta Ieven
May 8, 2019·PloS One·Daniel C MüllerAnders Johansson
Apr 10, 2020·Critical Reviews in Food Science and Nutrition·József LehelPéter Laczay
Feb 23, 2019·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·Ellen H A van den MunckhofMaurine A Leverstein-van Hall
Nov 12, 2020·BMC Infectious Diseases·Keke LiuYongqiang Jiang
Jan 31, 2020·Biosensors & Bioelectronics·Hanliang ZhuPavel Neužil
Sep 29, 2021·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Martin AgerhällDiana Berggren

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Methods Mentioned

BETA
PCR
Assay

Software Mentioned

Excel
SDS
GraphPad Prism
Primer Express

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