%0 Journal Article
%T Development of a Customized TaqMan£¿£¿ Array Card for Simultaneous Detection of 32 Respiratory Pathogens
%A Deborah Steensels
%A Isabel Montesinos
%A Katrien Lagrou
%A Kurt Beuselinck
%A Marie-Luce Delforge
%J -
%D 2018
%X Background: Syndromic testing with a rapid and complete diagnostic panel would be extremely useful for routine respiratory testing.
Objective: We developed and validated a real-time PCR-based Taqman array card (TAC) including several pathogens specific for immuno-compromised patients, using sequences designed and supported by the manufacturer.
Study design: Analytical validation was performed in 2 phases. Phase 1 testing was performed in 96-well plates and included efficiency, reactivity and sensitivity testing. The assays selected after phase 1 were validated in TAC format in phase 2 which included analytical sensitivity (LoD), reactivity, specificity and reproducibility testing. Clinical validation was performed in phase 3 by testing respiratory samples that were sent to the lab between December 2016 and May 2017.  Results: In total, 90 assays were tested in phase 1 of which 43 were retained for further validation in TAC format. Seven of the 43 assays did not meet the predefined criterion of efficiency ¡Ý 80%, but were accepted for further testing in phase 2 and 3 due to lack of alternatives. All 43 targets together detected 89% (481/539) of positives included in the reactivity testing in phase 1 and 98% (246/252) of positives in phase 2. All tested assays showed excellent analytical sensitivity, with LoD ranging from 1 to 100 copies/¦ÌL. An overall specificity of 99.96% was found. Reproducibility ranged from 76%-100%, with a mean of 91%. For the clinical validation, a total number of 428 samples were tested, with an overall positivity rate of 56.3% and a co-infection rate of 15.9%. 15 results (4% of total number of positives) were not confirmed and considered false positives. One result (0.2%) was considered false negative.
Conclusion: This syndromic panel was analytically and clinically validated and will be implemented for routine respiratory testing in the Erasme University Hospital in high risk patient populations.
%K TaqMan£¿£¿£¿£¿£¿£¿£¿£¿ array card
%K Real-time polymerase chain reaction/methods
%K Respiratory tract Infections/microbiology syndromic testing
%K Development and validation
%K Sensitivity
%K Specificity
%K Reproducibility
%K list of open access journals
%K open access
%K open access journals
%K open access publication
%K open access publisher
%K open access publishing
%K open access journal articles
%K imedpub
%K imedpub publishing
%K insight medical publishing
%K imedpub online
%U https://www.imedpub.com/articles/development-of-a-customized-taqman-array-card-for-simultaneous-detection-of-32-respiratory-pathogens.php?aid=21579