A loop-mediated isothermal amplification (LAMP) assay for the consensus detection of human pathogenic Campylobacter species
| Autor: | Elmer C. Bigley, Lisa M. Harrison, Mark K. Mammel, Uma S. Babu, Kelli L. Hiett, Kannan V. Balan |
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| Rok vydání: | 2020 |
| Předmět: |
Microbiology (medical)
DNA Bacterial Salmonella Loop-mediated isothermal amplification medicine.disease_cause Microbiology Campylobacter jejuni Sensitivity and Specificity 03 medical and health sciences RNA Ribosomal 16S Campylobacter Infections medicine Humans Shigella Helicobacter Molecular Biology 030304 developmental biology DNA Primers 0303 health sciences biology 030306 microbiology Campylobacter biology.organism_classification Molecular Diagnostic Techniques Arcobacter Campylobacter coli Nucleic Acid Amplification Techniques |
| Zdroj: | Journal of microbiological methods. 176 |
| ISSN: | 1872-8359 |
| Popis: | Most rapid identification methods for Campylobacter are designed to detect thermotolerant Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli). A growing number of thermosensitive Campylobacter species are now gaining recognition as emerging human pathogens. Methods are lacking for the rapid screening of these emerging species. Loop-mediated Isothermal Amplification (LAMP) is a nucleic acid amplification method that allows for the rapid and cost-effective detection of bacteria. Degenerate primers against the 16S rRNA sequences for C. jejuni, C. coli, C. lari, C. upsaliensis, C. ureolyticus, C. fetus, C. gracilis, C. rectus, and C. concisus were designed. Isothermal amplification was conducted using ATCC reference strains at 68 °C for 30 min using WarmStart® Colorimetric LAMP reagents. Positive reactions were indicated by a color change from pink to yellow; specificity to Campylobacter was confirmed using a restriction enzyme digest (RsaI). The developed LAMP reaction was specific for the reference strains, which was confirmed against an exclusivity panel that consisted of other enteric pathogens, including E. coli, Salmonella, Shigella, Helicobacter, and Arcobacter. This method was also evaluated for the detection of C. jejuni, C. coli, and C. lari in primary enrichment media from artificially contaminated fresh spinach samples. The LAMP method provides an option to rapidly screen for the presence of pathogenic Campylobacter spp. in field surveillance and trace-back analysis. |
| Databáze: | OpenAIRE |
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