Loop mediated isothermal amplification as a molecular diagnostic assay: Application and evaluation for detection of Enterohaemorrhagic Escherichia coli (O157:H7).
| Autor: | Yinur D; Department of Medical Biotechnology, Institute of Biotechnology, University of Gondar, Gondar, Ethiopia., Moges B; Department of Biotechnology, Debre Berhan University, Debre Berhan, Ethiopia., Hassen A; Department of Biotechnology, Ambo University, Ambo, Ethiopia., Tessema TS; Institute of Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia. |
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| Jazyk: | angličtina |
| Zdroj: | Practical laboratory medicine [Pract Lab Med] 2023 Sep 04; Vol. 37, pp. e00333. Date of Electronic Publication: 2023 Sep 04 (Print Publication: 2023). |
| DOI: | 10.1016/j.plabm.2023.e00333 |
| Abstrakt: | Purpose: This study aimed at evaluating the performance of the Loop Mediated Isothermal Amplification (LAMP) diagnostic test, which targets the putative Fimbria protein-encoding gene (Z3276) for rapid and specific detection of locally isolated enterohemorrhagic Escherichia coli (EHEC) O157:H7. Results: A total number of 40 locally available bacteria isolates and standard strains, among them 6 entrohemorrhagic (O157:H7) and 10 entropathogenic E. coli , 7 non diarrheic E. coli strains and 13 non entrohemorrhagic shiga toxic (stx) E. coli isolates as well as 4 pathogenic non E. coli species were used to optimize and evaluate the LAMP assay. The LAMP amplified DNA samples were visualized as turbid DNA both by naked eye and gel electrophoresis followed by staining. The assay had a sensitivity of 100% (6/6), a specificity of 97.05% (33/34), and an efficiency of 97.5% (39/40). The assay was also exhibited with 100% negative predicted value and 85.7% positive predicted value. The LAMP assay was also 10-fold more sensitive than the conventional PCR assay; sensitivity was determined by serial dilution. The results of LAMP and the PCR tests showed very high agreement (k = 0.97) in the detection of the bacteria studied. Conclusion: Compared with the performance of PCR and SMAC, LAMP assay was better in terms of efficiency, rapidity and cost-effectiveness, which can be used as a point-care diagnostic test in resource-limited laboratories. Competing Interests: The authors declare that they have no competing interests. (© 2023 The Authors.) |
| Databáze: | MEDLINE |
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