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Development of multilocus sequence typing (MLST) assay for Mycoplasma iowae
Mostafa Ghanem1, Mohamed El-Gazzar1
1Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH, 43210, USA.
Veterinary Microbiology
|October 25, 2016
Summary
A new multilocus sequence typing (MLST) assay was developed for identifying Mycoplasma iowae (MI) strains in turkeys. This reproducible method improves upon DNA fingerprinting for disease control.
Area of Science:
- Veterinary Microbiology
- Molecular Epidemiology
- Avian Pathology
Background:
- Mycoplasma iowae (MI) infection poses significant economic challenges in turkey farming.
- Current strain identification methods, like DNA fingerprinting, lack reproducibility and require pure cultures, which are difficult to obtain for avian mycoplasmas.
Purpose of the Study:
- To develop and validate the first multilocus sequence typing (MLST) assay for Mycoplasma iowae strain identification.
- To establish a reproducible and efficient molecular tool for tracking MI epidemiology.
Main Methods:
- Identification and analysis of 26 housekeeping gene loci from available Mycoplasma iowae genomes.
- Selection of six optimal loci (dppC, ulaA, valS, rpoC, leuS, kdpA) for the MLST assay.
- Typing of 47 MI samples using the developed MLST assay and subsequent phylogenetic analysis.
Main Results:
- The developed MLST assay successfully differentiated 47 MI samples into 23 unique sequence types using six loci.
- Even with only four loci, the assay distinguished 20 unique sequence types, demonstrating its discriminatory power.
- Phylogenetic analysis showed strong correlation between MLST data and geographical/temporal sample origins.
Conclusions:
- MLST is a highly reproducible molecular assay for Mycoplasma iowae identification and epidemiological surveillance.
- The assay enables direct identification from DNA samples, facilitating improved disease control and eradication strategies in turkeys.
