Isolation and molecular identification of Mycoplasma equigenitalium from equine genital tracts in northern India.
Abstract: Although Mycoplasma equigenitalium has been implicated in equine reproductive problems, its prevalence is largely unexplored due to the lack of specific diagnostic tests. To address this limitation, the authors developed and optimized species-specific primer pairs that target M. eguigenitalium rpoB (RNA polymerase B subunit) gene sequences. The specificity of the PCR assay developed in this study was determined using 12 field isolates including the type strain of M. equigenitalium and other Mycoplasma species. In the field study, a total of 122 mare and stallion samples comprising of 50 clinical and 72 random samples were subjected to species-specific PCR assay to detect M. equigenitalium in equine genital tracts. Mycoplasma equigenitalium (MEG) species-specific PCR detected 22.13% positive samples; however, only 9.01% of the samples were found to be positive using the conventional culture technique. The PCR established in this study could be used for rapid, specific and accurate diagnosis of M. equigenitalium strains. To the authors' knowledge, this is the first report addressing the development and evaluation of species-specific PCR to detect M. equigenitalium.
Publication Date: 2016-05-14 PubMed ID: 27175172PubMed Central: PMC4827683
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- Journal Article
Summary
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The research tackles the development of a species-specific PCR test to diagnose Mycoplasma equigenitalium, a bacteria causing reproductive problems in horses, with the goal to overcome current limitations in detecting the bacteria due to non-specific diagnostic tests.
Objective of the Research
- The study aimed to develop an optimized PCR (Polymerase Chain Reaction) assay targeting the rpoB gene sequences of Mycoplasma equigenitalium (MEG), an organism associated with reproductive issues in horses. By doing so, the researchers sought to improve the ability to diagnose this bacterial infection in horses by creating a more specific and accurate diagnostic test, given that the current tests available lack specificity.
Methods and Experimentation
- The researchers designed species-specific primer pairs to target the MEG’s rpoB gene sequences. A primer pair is a set of short nucleic acid sequences that provide a starting point for DNA synthesis in PCR testing.
- The specificity of the developed PCR assay was ascertained by using it on 12 different field isolates, which included the type strain of M. equigenitalium and other Mycoplasma types.
- A field study was also conducted where the developed PCR assay was used to test 122 equine genital samples (50 clinical and 72 random samples) for the presence of M. equigenitalium.
Results
- Applying the established PCR test, approximately 22.13% of the samples came back positive, whereas the conventional culture technique only detected M. equigenitalium in around 9.01% of the samples. The test developed in this study thus proved to be more accurate and specific in detecting MEG.
Conclusion
- The PCR test developed in the course of this study could aid in faster, more precise diagnoses of M. equigenitalium infections. Accordingly, this study sets a milestone as the first one that had addressed and successfully developed a species-specific PCR test for M. equigenitalium detection.
Cite This Article
APA
Nehra K, Rana R, Viswas KN, Arun TR, Singh VP, Singh AP, Prabhu SN.
(2016).
Isolation and molecular identification of Mycoplasma equigenitalium from equine genital tracts in northern India.
Iran J Vet Res, 16(2), 176-181.
Publication
Researcher Affiliations
- Graduated from Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India;
- Referral Lab on Mycoplasma, Division Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India;
- Division Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India;
- Graduated from Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India;
- Division of Bacteriology, Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India;
- Department of Microbiology, College of Veterinary & Animal Sciences (COVs&AH), Pt Deen Dayal Veterinary University (DUVASU), Mathura, 281001, Uttar Pradesh, India;
- Ph.D. Scholar in Veterinary Pathology, Division of Pathology, Indian Veterinary Research Institute, Izatnagar, 243122, Bareilly, Uttar Pradesh, India.
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Citations
This article has been cited 1 times.- Dawood A, Algharib SA, Zhao G, Zhu T, Qi M, Delai K, Hao Z, Marawan MA, Shirani I, Guo A. Mycoplasmas as Host Pantropic and Specific Pathogens: Clinical Implications, Gene Transfer, Virulence Factors, and Future Perspectives.. Front Cell Infect Microbiol 2022;12:855731.
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