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Ryo Harasawa - One of the best experts on this subject based on the ideXlab platform.
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Molecular detection of Mycoplasma haemomuris subspecies using dnaK-targeted real-time PCR with SYBR Green I and melting curve analysis
Molecular and Cellular Probes, 2016Co-Authors: Hiroshi Satoh, Daiki Imai, Shigeru Sato, Kazuhisa Furuhama, Ryo HarasawaAbstract:Abstract Hemoplasmas cause severe infections in mammals, but these pathogens are difficult to detect and identify at the species and subspecies level because of the need for time-consuming sequence based methods. Here, we used real-time PCR with SYBR Green I targeting of the dnaK gene followed by standard melting curve analysis to achieve rapid detection and differentiation of the Mycoplasma haemomuris subspecies ‘ Candidatus Mycoplasma haemomuris subsp. musculi’ and ‘ Candidatus M. haemomuris subsp. ratti’. The melting temperatures of the PCR products, 84.63 ± 0.14 °C for ‘ Candidatus M. haemomuris subsp. musculi’, and 80.72 ± 0.16 °C for ‘ Candidatus M. haemomuris subsp. ratti’, provided clear differentiation between them. Murine hemoplasma DNA samples, which were used as references, were confirmed for species by an analysis of 16S rRNA sequences. The protocol described herein provides a new rapid detection and identification method suitable for use with two recognized subspecies of M. haemomuris.
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Proposal for 'Candidatus Mycoplasma haemomuris subsp. musculi' in mice, and 'Candidatus Mycoplasma haemomuris subsp. ratti' in rats.
International Journal of Systematic and Evolutionary Microbiology, 2014Co-Authors: Ryo Harasawa, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Yasuko RikihisaAbstract:Mycoplasma haemomuris is causative of infectious anaemia or splenomegaly in rodents. We examined the nucleotide sequences of the non-ribosomal genes, rnpB and dnaK, in strains of the species M. haemomuris detected in small field mice and black rats. rnpB nucleotide sequences in strains of the species M. haemomuris isolated from small field mice and black rats had only 89 % sequence similarity, suggesting their separation into two distinct subgroups. dnaK had a nucleotide sequence similarity of 84 % between the subgroups. These results support the classification of M. haemomuris into two genetically distinct subgroups. Here we propose the establishment of these subgroups as ‘Candidatus Mycoplasma haemomuris subsp. musculi’, detected in small field mice (Apodemus argenteus), and ‘Candidatus Mycoplasma haemomuris subsp. ratti’, detected in black rats (Rattus rattus).
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Haemotropic Mycoplasma Infection Revealed by Real-Time PCR in Specific Pathogen-Free Rats
Bulletin of the Veterinary Institute in Pulawy, 2014Co-Authors: Hinako Sashida, Ryo Harasawa, Hiroshi Satoh, Toshihiro Ichijo, Kazuhisa FuruhamaAbstract:Abstract The presence of Mycoplasma haemomuris (haemoplasma) in blood samples collected from specific pathogen-free (SPF) laboratory rats bred in Japan was reported. Its presence was examined in Fischer 344, Sprague-Dawley (SD), and Wistar rat strains of both sexes by real-time PCR. All strains were positive for M. haemomuris infection. The 16S rRNA gene of M. haemomuris strain detected in the animals was amplified using end-point PCR. Only the entire nucleotide sequence of 16S rRNA gene of a Mycoplasma strain detected in SD rats was determined and compared to those of other haemoplasmas. Our investigations suggest a wide M. haemomuris infection among the SPF rats purchased from commercial breeders in Japan.
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Two clusters among Mycoplasma haemomuris strains, defined by the 16S-23S rRNA intergenic transcribed spacer sequences.
Journal of Veterinary Medical Science, 2013Co-Authors: Hinako Sashida, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Fumina Sasaoka, Jin Suzuki, Masatoshi Fujihara, Kazuya Nagai, Ryo HarasawaAbstract:Mycoplasma haemomuris is a causative organism of infectious anemia or splenomegaly in rodents. Here, we report two distinct genetic groups among M. haemomuris strains detected from rats and mice, respectively, by examining the nucleotide sequences of the 16S-23S rRNA intergenic transcribed spacer region that has been shown to be a stable genetic marker for Mycoplasma species. Our results may reveal host-tropism of each cluster of M. haemomuris strains, and suggest an idea to distinguish M. haemomuris into two different genetic clusters.
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Characteristics of the 16S-23S rRNA intergenic spacer region of Mycoplasma haemomuris, previously classified as 'Haemobartonella muris'.
Journal of Veterinary Medical Science, 2002Co-Authors: Ryo Harasawa, Makoto Kawahara, Yasuko RikihisaAbstract:The intergenic spacer region between the 16S and 23S rRNA genes of Mycoplasma haemomuris, previously classified as `Haemobartonella muris', was amplified by PCR and sequenced for analysis of the primary and secondary structures of the RNA transcript. The spacer region consisted of 219 base-pairs and lacked the spacer tRNA gene. A hypothetical secondary structure predicted in the RNA transcript of the spacer region was tentatively assigned boxA and boxB loci peculiar to the members of the Mycoplasma. Mycoplasma haemomuris and the other species of the genus Mycoplasma are consistent with these characteristics of the spacer region.
Yasuko Rikihisa - One of the best experts on this subject based on the ideXlab platform.
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Proposal for 'Candidatus Mycoplasma haemomuris subsp. musculi' in mice, and 'Candidatus Mycoplasma haemomuris subsp. ratti' in rats.
International Journal of Systematic and Evolutionary Microbiology, 2014Co-Authors: Ryo Harasawa, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Yasuko RikihisaAbstract:Mycoplasma haemomuris is causative of infectious anaemia or splenomegaly in rodents. We examined the nucleotide sequences of the non-ribosomal genes, rnpB and dnaK, in strains of the species M. haemomuris detected in small field mice and black rats. rnpB nucleotide sequences in strains of the species M. haemomuris isolated from small field mice and black rats had only 89 % sequence similarity, suggesting their separation into two distinct subgroups. dnaK had a nucleotide sequence similarity of 84 % between the subgroups. These results support the classification of M. haemomuris into two genetically distinct subgroups. Here we propose the establishment of these subgroups as ‘Candidatus Mycoplasma haemomuris subsp. musculi’, detected in small field mice (Apodemus argenteus), and ‘Candidatus Mycoplasma haemomuris subsp. ratti’, detected in black rats (Rattus rattus).
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Revision of haemotrophic Mycoplasma species names.
International Journal of Systematic and Evolutionary Microbiology, 2002Co-Authors: Harold Neimark, Yasuko Rikihisa, Karl-erik Johansson, Joseph G. TullyAbstract:The recently proposed transfer of four rickettsias from the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with the Candidatus status is herein revised. This is because the Candidatus designation is for new, incompletely described taxa, in order to give them a provisional status. Thus, 'Candidatus Mycoplasma haemofelis' is revised to Mycoplasma haemofelis comb. nov., nom. nov., 'Candidatus Mycoplasma haemomuris' is revised to Mycoplasma haemomuris comb. nov., nom. nov., 'Candidatus Mycoplasma haemosuis' is revised to Mycoplasma haemosuis comb. nov., nom. nov. and 'Candidatus Mycoplasma wenyonii' is revised to Mycoplasma wenyonii comb. nov.
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Characteristics of the 16S-23S rRNA intergenic spacer region of Mycoplasma haemomuris, previously classified as 'Haemobartonella muris'.
Journal of Veterinary Medical Science, 2002Co-Authors: Ryo Harasawa, Makoto Kawahara, Yasuko RikihisaAbstract:The intergenic spacer region between the 16S and 23S rRNA genes of Mycoplasma haemomuris, previously classified as `Haemobartonella muris', was amplified by PCR and sequenced for analysis of the primary and secondary structures of the RNA transcript. The spacer region consisted of 219 base-pairs and lacked the spacer tRNA gene. A hypothetical secondary structure predicted in the RNA transcript of the spacer region was tentatively assigned boxA and boxB loci peculiar to the members of the Mycoplasma. Mycoplasma haemomuris and the other species of the genus Mycoplasma are consistent with these characteristics of the spacer region.
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Evaluation of sensitivity and specificity of a Mycoplasma haemomuris-specific polymerase chain reaction test.
Comparative medicine, 2002Co-Authors: Chunbin Zhang, Yasuko RikihisaAbstract:BACKGROUND AND PURPOSE Mycoplasma haemomuris, a small pleomorphic bacterium parasitic of red blood cells, often causes chronic and subclinical infection of rodents. Mycoplasma haemomuris is uncultivable, and a serologic testing method is not readily available. The purpose of the study reported here was to develop a sensitive and specific polymerase chain reaction (PCR) test for detection of M. haemomuris in blood samples. METHODS On the basis of the regions of the M. haemomuris 16S rRNA gene most divergent from corresponding regions of related bacteria, M. haemomuris-specific primers were designed so that these primers could selectively amplify M. haemomuris DNA. A PCR test was performed, using blood samples from BALB/c mice infected with M. haemomuris strains TR 8564, TR 8563, and TR 8556. RESULTS Use of the PCR test enabled detection of M. haemomuris DNA in a minimum of 0.0001 microl of infected mouse blood. The test also was specific for M. haemomuris and did not amplify closely related species, such as M. haemofelis, M. haemosuis, M. orale, or Anaplasma marginale. CONCLUSION This method is sensitive and specific for detection of M. haemomuris.
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Proposal to transfer some members of the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with descriptions of 'Candidatus Mycoplasma haemofelis', 'Candidatus Mycoplasma haemomuris', 'Candidatus Mycoplasma haemosuis' and 'Candidatus M
International Journal of Systematic and Evolutionary Microbiology, 2001Co-Authors: Harold Neimark, Yasuko Rikihisa, Karl-erik Johansson, Joseph G. TullyAbstract:Cell-wall-less uncultivated parasitic bacteria that attach to the surface of host erythrocytes currently are classified in the order Rickettsiales, family Anaplasmataceae, in the genera Haemobartonella and Eperythrozoon. Recently 16S rRNA gene sequences have been determined for four of these species: Haemobartonella felis and Haemobartonella muris and Eperythrozoon suis and Eperythrozoon wenyonii. Phylogenetic analysis of these sequence data shows that these haemotrophic bacteria are closely related to species in the genus Mycoplasma (class Mollicutes). These haemotrophic bacteria form a new phylogenetic cluster within the so-called pneumoniae group of Mycoplasma and share properties with one another as well as with other members of the pneumoniae group. These studies clearly indicate that the classification of these taxa should be changed to reflect their phylogenetic affiliation and the following is proposed: (i) that Haemobartonella felis and Haemobartonella muris should be transferred to the genus Mycoplasma as 'Candidatus Mycoplasma haemofelis' and 'Candidatus Mycoplasma haemomuris' and (ii) that Eperythrozoon suis and Eperythrozoon wenyonii should be transferred to the genus Mycoplasma as 'Candidatus Mycoplasma haemosuis' and 'Candidatus Mycoplasma wenyonii'. The former Haemobartonella and Eperythrozoon species described here represent a new group of parasitic Mycoplasmas that possess a pathogenic capacity previously unrecognized among the mollicutes. These haemotrophic Mycoplasmas have been given the trivial name haemoplasmas. These results call into question the affiliation of the remaining officially named species of Haemobartonella and Eperythrozoon which should be considered species of uncertain affiliation pending the resolution of their phylogenetic status.
Hinako Sashida - One of the best experts on this subject based on the ideXlab platform.
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Haemotropic Mycoplasma Infection Revealed by Real-Time PCR in Specific Pathogen-Free Rats
Bulletin of the Veterinary Institute in Pulawy, 2014Co-Authors: Hinako Sashida, Ryo Harasawa, Hiroshi Satoh, Toshihiro Ichijo, Kazuhisa FuruhamaAbstract:Abstract The presence of Mycoplasma haemomuris (haemoplasma) in blood samples collected from specific pathogen-free (SPF) laboratory rats bred in Japan was reported. Its presence was examined in Fischer 344, Sprague-Dawley (SD), and Wistar rat strains of both sexes by real-time PCR. All strains were positive for M. haemomuris infection. The 16S rRNA gene of M. haemomuris strain detected in the animals was amplified using end-point PCR. Only the entire nucleotide sequence of 16S rRNA gene of a Mycoplasma strain detected in SD rats was determined and compared to those of other haemoplasmas. Our investigations suggest a wide M. haemomuris infection among the SPF rats purchased from commercial breeders in Japan.
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Two clusters among Mycoplasma haemomuris strains, defined by the 16S-23S rRNA intergenic transcribed spacer sequences.
Journal of Veterinary Medical Science, 2013Co-Authors: Hinako Sashida, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Fumina Sasaoka, Jin Suzuki, Masatoshi Fujihara, Kazuya Nagai, Ryo HarasawaAbstract:Mycoplasma haemomuris is a causative organism of infectious anemia or splenomegaly in rodents. Here, we report two distinct genetic groups among M. haemomuris strains detected from rats and mice, respectively, by examining the nucleotide sequences of the 16S-23S rRNA intergenic transcribed spacer region that has been shown to be a stable genetic marker for Mycoplasma species. Our results may reveal host-tropism of each cluster of M. haemomuris strains, and suggest an idea to distinguish M. haemomuris into two different genetic clusters.
Kazuhisa Furuhama - One of the best experts on this subject based on the ideXlab platform.
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Molecular detection of Mycoplasma haemomuris subspecies using dnaK-targeted real-time PCR with SYBR Green I and melting curve analysis
Molecular and Cellular Probes, 2016Co-Authors: Hiroshi Satoh, Daiki Imai, Shigeru Sato, Kazuhisa Furuhama, Ryo HarasawaAbstract:Abstract Hemoplasmas cause severe infections in mammals, but these pathogens are difficult to detect and identify at the species and subspecies level because of the need for time-consuming sequence based methods. Here, we used real-time PCR with SYBR Green I targeting of the dnaK gene followed by standard melting curve analysis to achieve rapid detection and differentiation of the Mycoplasma haemomuris subspecies ‘ Candidatus Mycoplasma haemomuris subsp. musculi’ and ‘ Candidatus M. haemomuris subsp. ratti’. The melting temperatures of the PCR products, 84.63 ± 0.14 °C for ‘ Candidatus M. haemomuris subsp. musculi’, and 80.72 ± 0.16 °C for ‘ Candidatus M. haemomuris subsp. ratti’, provided clear differentiation between them. Murine hemoplasma DNA samples, which were used as references, were confirmed for species by an analysis of 16S rRNA sequences. The protocol described herein provides a new rapid detection and identification method suitable for use with two recognized subspecies of M. haemomuris.
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Haemotropic Mycoplasma Infection Revealed by Real-Time PCR in Specific Pathogen-Free Rats
Bulletin of the Veterinary Institute in Pulawy, 2014Co-Authors: Hinako Sashida, Ryo Harasawa, Hiroshi Satoh, Toshihiro Ichijo, Kazuhisa FuruhamaAbstract:Abstract The presence of Mycoplasma haemomuris (haemoplasma) in blood samples collected from specific pathogen-free (SPF) laboratory rats bred in Japan was reported. Its presence was examined in Fischer 344, Sprague-Dawley (SD), and Wistar rat strains of both sexes by real-time PCR. All strains were positive for M. haemomuris infection. The 16S rRNA gene of M. haemomuris strain detected in the animals was amplified using end-point PCR. Only the entire nucleotide sequence of 16S rRNA gene of a Mycoplasma strain detected in SD rats was determined and compared to those of other haemoplasmas. Our investigations suggest a wide M. haemomuris infection among the SPF rats purchased from commercial breeders in Japan.
Hiromi Fujita - One of the best experts on this subject based on the ideXlab platform.
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Proposal for 'Candidatus Mycoplasma haemomuris subsp. musculi' in mice, and 'Candidatus Mycoplasma haemomuris subsp. ratti' in rats.
International Journal of Systematic and Evolutionary Microbiology, 2014Co-Authors: Ryo Harasawa, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Yasuko RikihisaAbstract:Mycoplasma haemomuris is causative of infectious anaemia or splenomegaly in rodents. We examined the nucleotide sequences of the non-ribosomal genes, rnpB and dnaK, in strains of the species M. haemomuris detected in small field mice and black rats. rnpB nucleotide sequences in strains of the species M. haemomuris isolated from small field mice and black rats had only 89 % sequence similarity, suggesting their separation into two distinct subgroups. dnaK had a nucleotide sequence similarity of 84 % between the subgroups. These results support the classification of M. haemomuris into two genetically distinct subgroups. Here we propose the establishment of these subgroups as ‘Candidatus Mycoplasma haemomuris subsp. musculi’, detected in small field mice (Apodemus argenteus), and ‘Candidatus Mycoplasma haemomuris subsp. ratti’, detected in black rats (Rattus rattus).
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Two clusters among Mycoplasma haemomuris strains, defined by the 16S-23S rRNA intergenic transcribed spacer sequences.
Journal of Veterinary Medical Science, 2013Co-Authors: Hinako Sashida, Hiromi Fujita, Teruki Kadosaka, Shuji Ando, Fumina Sasaoka, Jin Suzuki, Masatoshi Fujihara, Kazuya Nagai, Ryo HarasawaAbstract:Mycoplasma haemomuris is a causative organism of infectious anemia or splenomegaly in rodents. Here, we report two distinct genetic groups among M. haemomuris strains detected from rats and mice, respectively, by examining the nucleotide sequences of the 16S-23S rRNA intergenic transcribed spacer region that has been shown to be a stable genetic marker for Mycoplasma species. Our results may reveal host-tropism of each cluster of M. haemomuris strains, and suggest an idea to distinguish M. haemomuris into two different genetic clusters.
