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Development of RFLP-PCR and simple multiplex PCR assays for detection and differentiation of two species of hemotropic mycoplasmas in naturally infected dogs

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Abstract

Canine hemotropic mycoplasmas occur worldwide and are caused by the uncultivable hemotrophic bacteria; Mycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp). In this study, the occurrence and differentiation of both causative agents in dogs that originated from southern Iran were investigated by employing restriction fragment length polymorphism (RFLP)-PCR and simple multiplex PCR methods based on RNase P RNA (rnpB) and 16S rDNA markers, respectively. The results showed that the prevalence of Mhc and CMhp among 100 examined samples were 6.0 and 4.0 %, respectively. Both PCR assays provide valuable molecular tools for the differentiation of CMhp and Mhc DNA in canine blood samples compared to 16S rDNA sequencing results. Also, sequence analysis of the genes (rnpB and 16S rDNA) and subsequent phylogenetic analysis were performed for both hemoplasmas in the Iranian dogs’ population. Comparison of the Iranian sequences with those for other isolates of canine hemoplasmas deposited in the GenBank database revealed a higher interspecific variability for rnpB marker (up to 31.22 %) compared to 16S rDNA sequence (up to 17 %).

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Authors and Affiliations

  1. Department of Clinical Studies, School of Veterinary Medicine, Shiraz University, P.O. Box 1731, 71345, Shiraz, Iran

    Hassan Sharifiyazdi, Mohammad Abbaszadeh Hasiri & Mahshid Radmanesh

Authors
  1. Hassan Sharifiyazdi
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  2. Mohammad Abbaszadeh Hasiri
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  3. Mahshid Radmanesh
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Corresponding author

Correspondence to Hassan Sharifiyazdi.

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Ethical approval

This experiment was performed under the approval of the state committee on animal ethics, Shiraz University, Shiraz, Iran. In addition, the recommendations of European Council Directive (86/609/EC) of November 24, 1986, regarding the protection of animals used for experimental purposes, were also considered.

Funding

This study was funded by Research Council of Shiraz University and School of Veterinary Medicine, Shiraz University (Grant No. 87-GR-VT-47).

Conflict of interest

The authors declare that they have no conflict of interest.

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Sharifiyazdi, H., Abbaszadeh Hasiri, M. & Radmanesh, M. Development of RFLP-PCR and simple multiplex PCR assays for detection and differentiation of two species of hemotropic mycoplasmas in naturally infected dogs. Comp Clin Pathol 25, 847–853 (2016). https://doi.org/10.1007/s00580-016-2272-7

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  • DOI: https://doi.org/10.1007/s00580-016-2272-7

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