Comparative analysis of two broad-range PCR assays for pathogen detection in positive-blood-culture bottles: PCR-high-resolution melting analysis versus PCR-mass spectrometry
| Autor: | Samuel Yang, Karen C. Carroll, Helen Won, Yu Hsiang Hsieh, Heather E. Matthews, Rangarajan Sampath, Richard E. Rothman, Alexander Kecojevic, Lawrence B. Blyn, Donna Toleno, Charlotte A. Gaydos, Stephen Peterson, Kevin Jeng, Justin Hardick, Billy Masek |
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| Rok vydání: | 2012 |
| Předmět: |
Microbiology (medical)
Microbiological Techniques Concordance Pcr assay Pilot Projects Biology Mass spectrometry Nucleic Acid Denaturation Polymerase Chain Reaction Sensitivity and Specificity High Resolution Melt Melting curve analysis Mass Spectrometry law.invention law Sepsis medicine Humans Transition Temperature Blood culture Polymerase chain reaction Retrospective Studies medicine.diagnostic_test Bacteriology Amplicon Molecular biology Blood Molecular Diagnostic Techniques |
| Zdroj: | Journal of clinical microbiology. 50(10) |
| ISSN: | 1098-660X |
| Popis: | Detection of pathogens in bloodstream infections is important for directing antimicrobial treatment, but current culture-based approaches can be problematic. Broad-range PCR assays which target conserved genomic motifs for postamplification amplicon analysis permit detection of sepsis-causing pathogens. Comparison of different broad-range assays is important for informing future implementation strategies. In this study, we compared positive-blood-culture bottles processed by PCR coupled to high-resolution melting curve analysis (PCR/HRMA) and PCR coupled to electrospray ionization-mass spectrometry (PCR/ESI-MS) to microbiology culture results. Genus-level concordance was 90% (confidence interval [CI], 80 to 96%) for PCR/HRMA and 94% (CI, 85 to 98%) for PCR/ESI-MS. Species-level concordance was 90% (CI, 80 to 96%) for PCR/HRMA and 86% (CI, 75 to 93%) for PCR/ESI-MS. Unlike PCR/HRMA, PCR/ESI-MS was able to resolve polymicrobial samples. Our results demonstrated that the two assays have similar overall concordance rates but may have different roles as potential adjunctive tests with standard blood culture, since each method has different capabilities, advantages, and disadvantages. |
| Databáze: | OpenAIRE |
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