Rapid detection of nusG and fadA in Fusobacterium nucleatum by loop-mediated isothermal amplification
| Autor: | Zhan Yang, Simo Huang, Liuyu Huang, Anheng Liu, Wei Liu, Dayang Zou, Derong Dong |
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| Rok vydání: | 2016 |
| Předmět: |
Adult
Male 0301 basic medicine Microbiology (medical) Time Factors 030106 microbiology Loop-mediated isothermal amplification Biology Sensitivity and Specificity Microbiology Rapid detection Virulence factor 03 medical and health sciences Bacterial Proteins stomatognathic system Periodontal disease Humans Gene Pathogen Aged DNA Primers Fusobacterium nucleatum General Medicine Middle Aged biology.organism_classification stomatognathic diseases 030104 developmental biology Molecular Diagnostic Techniques Fusobacterium Infections Female Primer (molecular biology) Nucleic Acid Amplification Techniques |
| Zdroj: | Journal of Medical Microbiology. 65:760-769 |
| ISSN: | 1473-5644 0022-2615 |
| DOI: | 10.1099/jmm.0.000300 |
| Popis: | Fusobacterium nucleatum is associated with various human diseases such as periodontal disease and colorectal cancer (CRC); thus, F. nucleatum detection might serve as a novel diagnostic tool. Here, we describe the development of a sensitive and rapid molecular method for detecting two F. nucleatum genes: the highly conserved nusG and fadA, which encode a critical host colonization factor. Loop-mediated isothermal amplification (LAMP) primer sets for the rapid detection of nusG and fadA were designed and optimized. The nusG primers yielded consistent negative results for 20 non-F. nucleatum bacterial strains, confirming the high specificity of the primers. LAMP reaction primer sensitivity was determined, and its detection rate in comparison to conventional PCR was assessed using 57 clinical stool samples. The LAMP detection limit for nusG and fadA was 22.5 and 0.225 pg µl-1, respectively, indicating that the sensitivity of this method was 10-fold higher than that of conventional PCR. These results suggest that the LAMP technique is able to effectively identify F. nucleatum via nusG as well as detect its virulence factor. To the best of our knowledge, this study is the first to report the application of LAMP for the detection of nusG and fadA in F. nucleatum. The LAMP method constitutes a sensitive and specific visual assay for the rapid detection of the pathogen F. nucleatum. |
| Databáze: | OpenAIRE |
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