Back to Search
Start Over
A novel real-time PCR assay panel for detection of common respiratory pathogens in a convenient, strip-tube array format
- Source :
- Journal of Virological Methods
- Publication Year :
- 2018
- Publisher :
- Cold Spring Harbor Laboratory, 2018.
-
Abstract
- Highlights • Respanel is a laboratory-developed PCR test panel for common respiratory pathogens. • Strip-tube array-based assay, designed for convenience and minimal hands-on time. • Specificity and accuracy of Respanel is ≥95% against two commercial assays. • Sensitivity of Respanel is 94% and 88% against two commercial assays.<br />Commercial multiplex assays, built on different chemistries and platforms are widely available for simultaneous detection of pathogens that cause respiratory infections. However, these tests are often difficult to implement in a resource limited setting because of high cost. In this study, we developed and validated a method for simultaneous testing of common respiratory pathogens (Respanel) by real-time PCR in a convenient, strip-tube array format. Primers and probes for sixteen PCR assays were selected from the literature or newly designed. Following optimization of individual PCR assays, strip-tube arrays were prepared by dispensing primer-probe mixes (PPM) into two sets of 8-tube strips. Nucleic acid extracts from specimens were mixed with PCR master mix, and dispensed column-wise into 2 × 8-wells of a 96-well plate. PPMs from strip-tubes were then added to the wells using a multichannel pipette for real-time PCR. Individual PCR assays were optimized using previously known specimens (n = 394) with 91%–100% concordance with culture, DFA or PCR results. Respanel was then tested in a routine manner at two different sites using specimens (n = 147) previously tested by Qiagen Resplex I&II or Fast-Track Diagnostics Respiratory Pathogens 21 assays. The sensitivity, specificity and accuracy of Respanel were 94%, 95% and 95%, respectively, against Resplex and 88%, 100% and 99%, respectively, against FTDRP21. Respanel detected more pathogens (p
- Subjects :
- 0301 basic medicine
Chromatography
Respiratory pathogens
030106 microbiology
Pcr assay
Pipette
Biology
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Article
03 medical and health sciences
030104 developmental biology
Real-time polymerase chain reaction
Molecular Diagnostic Techniques
Virology
Validation
Commercial multiplex assays
Costs and Cost Analysis
Multiplex
Multiplex Polymerase Chain Reaction
Respiratory Tract Infections
Limited resources
Real-time PCR
Laboratory-developed tests
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Journal of Virological Methods
- Accession number :
- edsair.doi.dedup.....706765c1c5b4209e70fbf68c751efff0