دورية أكاديمية
Verification of monoplex and multiplex linear-after-the-exponential PCR gene-specific sepsis assays using clinical isolates.
| العنوان: | Verification of monoplex and multiplex linear-after-the-exponential PCR gene-specific sepsis assays using clinical isolates. |
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| المؤلفون: | Gentile NL; Point-of-Care Testing Center for Teaching and Research (POCT•CTR), Pathology and Laboratory Medicine, School of Medicine, University of California, Davis, CA 95616, USA. nlgentile@ucdavis.edu, Dillier AM, Williams GV, Ackers J, Reis AH Jr, Rice LM, Wangh LJ, Czajka JW, Kost GJ |
| المصدر: | Journal of applied microbiology [J Appl Microbiol] 2013 Feb; Vol. 114 (2), pp. 586-94. Date of Electronic Publication: 2013 Jan 07. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: Oxford University Press Country of Publication: England NLM ID: 9706280 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1365-2672 (Electronic) Linking ISSN: 13645072 NLM ISO Abbreviation: J Appl Microbiol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2022- : Oxford : Oxford University Press Original Publication: Oxford : Published for the Society for Applied Bacteriology by Blackwell Science, c1997- |
| مواضيع طبية MeSH: | Bacteria/*isolation & purification , Polymerase Chain Reaction/*methods , Sepsis/*microbiology, Acinetobacter baumannii/genetics ; Acinetobacter baumannii/isolation & purification ; Candida/genetics ; Candida/isolation & purification ; DNA, Bacterial/analysis ; DNA, Bacterial/isolation & purification ; DNA, Fungal/analysis ; DNA, Fungal/isolation & purification ; Enterobacter/genetics ; Enterobacter/isolation & purification ; Enterococcus/genetics ; Enterococcus/isolation & purification ; Genes, Bacterial ; Genes, Fungal ; Humans ; Klebsiella/genetics ; Klebsiella/isolation & purification ; Multiplex Polymerase Chain Reaction/methods ; Pseudomonas aeruginosa/genetics ; Pseudomonas aeruginosa/isolation & purification ; Sensitivity and Specificity ; Staphylococcus/genetics ; Staphylococcus/isolation & purification ; Staphylococcus aureus/genetics ; Staphylococcus aureus/isolation & purification |
| مستخلص: | Aims: To verify monoplex and multiplex gene-specific linear-after-the-exponential polymerase chain reaction (LATE-PCR) assays for identifying 17 microbial pathogens (i.e., Klebsiella sp., Acinetobacter baumannii, Staphylococcus aureus, Enterobacter sp., Pseudomonas aeruginosa, coagulase negative staphylococci, Enterococcus sp., Candida sp.) commonly associated with septicaemia using clinical isolates. Methods and Results: Clinical isolates of each target pathogen were collected from the University of California, Davis Medical Center (UCDMC) microbiology laboratory. Five microlitres (μl) of each culture suspension (1 × 10(8) CFU ml(-1) ) were added to 20 μl of monoplex mastermix. DNA extracted from clinical isolates was tested in multiplex. Monoplex assays demonstrated 100% sensitivity at this input level, except Enterobacter cloacae (2.7%), Ac. baumannii (57%) and Ps. aeruginosa (97.8%). All clinical isolates were positive in multiplex, with the exception of two Ac. baumannii, two Klebsiella oxytoca and two Candida parapsilosis isolates. Conclusions: Sixteen pathogens can be identified by monoplex LATE-PCR assays with sensitivities ≥ 97.8%. The multiplex assay demonstrated 91.4% sensitivity when tested with DNA extracted from 70 different target strains. Significance and Impact of the Study: This study demonstrates the potential of LATE-PCR to serve as an adjunct to culture if the reagents are optimized for sensitivity. Results warrant further testing through analytical and clinical validation of the multiplex assay. (© 2012 The Society for Applied Microbiology.) |
| معلومات مُعتمدة: | RC1EB010643 United States EB NIBIB NIH HHS |
| المشرفين على المادة: | 0 (DNA, Bacterial) 0 (DNA, Fungal) |
| تواريخ الأحداث: | Date Created: 20121110 Date Completed: 20131209 Latest Revision: 20181202 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1111/jam.12062 |
| PMID: | 23136961 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1365-2672 |
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| DOI: | 10.1111/jam.12062 |