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Applied and Environmental Microbiology Apr 1993DNA of 10 lines of rice yellow dwarf (RYD) mycoplasmalike organisms (MLOs) from Japan, the Phillippines, and Thailand hybridized with four probes containing chromosomal...
DNA of 10 lines of rice yellow dwarf (RYD) mycoplasmalike organisms (MLOs) from Japan, the Phillippines, and Thailand hybridized with four probes containing chromosomal and six probes containing extrachromosomal DNA of a Tochigi (Japan) line of RYD MLO. One chromosomal probe (RYD9) and all six extrachromosomal probes hybridized with various other MLOs (sugarcane white leaf, onion yellows, cineraria witches'-broom, Japanese hornwort witches'-broom, water dropwort wiches'-broom, gentian witches'-broom, udo dwarf, tsuwabuki witches'-broom, pelargonium witches's-broom, peach western-X, and pear decline). DNA from the culturable mollicutes Spiroplasma kunkelii, Spiroplasma citri, Mycoplasma hominis, and Mycoplasma orale did not hybridize with RYD MLO probes. The extrachromosomal DNAs hybridizing with the probes showed variations in electrophoretic behavior.
Topics: Base Sequence; Chromosomes; DNA Probes; DNA, Bacterial; Molecular Sequence Data; Mycoplasma; Oryza; Plasmids; Tenericutes
PubMed: 8489230
DOI: 10.1128/aem.59.4.1206-1212.1993 -
Transfusion Medicine and Hemotherapy :... Feb 2014Contamination of cell culture and biological material by mollicute species is an important safety issue and requires testing. We have developed a singletube real-time...
BACKGROUND
Contamination of cell culture and biological material by mollicute species is an important safety issue and requires testing. We have developed a singletube real-time polymerase chain reaction (PCR) assay for rapid detection of Mollicutes species stipulated by the European Pharmacopeia.
METHODS
Primers and TaqMan probes (FAM-labeled) were deduced from 16S rDNA sequence alignment of 18 mollicutes species. A synthetic internal control (IC) DNA and an IC-specific TaqMan probe (VIC-labeled) were included. The analytical sensitivity of the assay was determined on DNA dilutions from 12 mollicute strains. Specificity was proven by the use of DNA from other bacteria.
RESULTS
Analytical sensitivities of the PCR assay were in the range of 405-2,431 genomes/ml for 11 of the 12 tested mollicute DNA samples. The lowest sensitivity was found for Ureaplasma urealyticum (19,239 genomes/ml). Negative results for DNA samples from 3 different ubiquitous bacteria demonstrated the specificity of the PCR assay for Mollicutes. Direct testing of cell culture supernatants spiked with Mycoplasma orale revealed similar sensitivity compared to isolated DNA.
CONCLUSIONS
Our single-tube real-time PCR assay with internal reaction control enables rapid and specific detection of mollicute contaminants. The test protocol is suitable for routine quality control of cell therapeutics.
PubMed: 24659951
DOI: 10.1159/000357096 -
Journal of Bacteriology Nov 1986Mycoplasma salivarium produced citrulline, ammonia, and ATP from N-benzoylglycyl-L-arginine. The activity was inhibited by EDTA and was therefore concluded to be due to...
Mycoplasma salivarium produced citrulline, ammonia, and ATP from N-benzoylglycyl-L-arginine. The activity was inhibited by EDTA and was therefore concluded to be due to an arginine-specific carboxypeptidase. The activity was also found to exist in M. orale, M. buccale, M. faucium, and M. hominis.
Topics: Adenosine Triphosphate; Ammonia; Arginine; Carboxypeptidases; Citrulline; Humans; Lysine Carboxypeptidase; Mycoplasma; Mycoplasma pneumoniae
PubMed: 3096956
DOI: 10.1128/jb.168.2.1045-1047.1986 -
Microbiology and Immunology 1982The cilia-stopping effect of mycoplasmas of human and various animal origin in mouse and chicken tracheal organ cultures was studied. From the results in mouse tracheal... (Comparative Study)
Comparative Study
The cilia-stopping effect of mycoplasmas of human and various animal origin in mouse and chicken tracheal organ cultures was studied. From the results in mouse tracheal organ cultures, the mycoplasma strains tested were divided into three groups: Mycoplasma pulmonis m53, M pulmonis JB, M. pulmonis OK, M. mycoides subsp. Mycoides PGl and M. Gallisepticum S6 showed a strong cilia-stopping effect; M. pulmonis PG22, M. mycoides subsp. capri PG3, M. meleagridis 19729, M. neurolyticum Type A and M. arthritidis PG6 showed a mild effect; and M. pneumoniae FH, M. salivarium Hup, M. hominis type 1-C and M. orale N-C human origin and Acholeplasma laidlawii PG8 showed a weak effect. On the other hand, in chicken tracheal organ cultures, only M. gallisepticum S6 showed a strong effect, M. meleagridis 19729 was affected to a lesser degree, and the other mycoplasma strains showed a weak or no effect. The results indicate that some murine and poultry mycoplasmas showed a cilia-stopping tendency in mouse and chicken tracheal organ cultures, respectively, while human mycoplasmas showed weak or little effect in both organ cultures. In mouse tracheal organ cultures, M. pulmonis m53 treated with heat, trypsin or formaldehyde, and the sterile filtrate of an m53 broth culture showed no cilia-stopping effect. The relationship of the pathogenicity of mycoplasmas for their natural hosts to that for cultured respiratory cells is discussed.
Topics: Animals; Chickens; Cilia; Humans; Mice; Microscopy, Electron, Scanning; Movement; Mycoplasma; Organ Culture Techniques; Trachea
PubMed: 7087799
DOI: 10.1111/j.1348-0421.1982.tb00148.x -
Journal of Bacteriology Nov 1977In this report we present the first description of the isolation and partial characterization of the deoxyribonucleic acid (DNA) polymerase activity from two species of...
In this report we present the first description of the isolation and partial characterization of the deoxyribonucleic acid (DNA) polymerase activity from two species of Mycoplasmatales, Mycoplasma orale type 1 and M. hyorhinis. We have identified only a single DNA polymerase species in the mycoplasma crude extracts, and the enzymes from the two organisms are very similar in their structural and enzymatic properties. The purified polymerase from each source has a specific activity of greater than 50,000 U/mg of protein, a sedimentation coefficient of 5.6s, and an estimated molecular weight by gel filtration of 130,000. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the most highly purified M. orale fraction contains a single major protein band of 130,000 daltons, which we believe may represent the polymerase protein. The enzymes are most reactive with gapped (activated) DNA and show a marked preference for this primer template over oligodeoxyribonucleotide-initiated homoribo- or homodeoxyribo-polymers. The most purified preparations are devoid of contaminating endonuclease activity and also appear to lack associated 5' leads to 3'- or 3' leads to 5'-exonuclease activities, as determined by highly sensitive assays. The absence of the 3' leads to 5'-exonuclease is particularly remarkable in that this activity is essentially ubiquitous among the DNA polymerases that have thus far been characterized from procaryotes.
Topics: Bacterial Proteins; Cell-Free System; DNA; DNA-Directed DNA Polymerase; Mercaptoethanol; Molecular Weight; Mycoplasma; Poly dA-dT; Pyrimidine Dimers; Sulfhydryl Reagents; Templates, Genetic
PubMed: 914780
DOI: 10.1128/jb.132.2.641-649.1977 -
Applied and Environmental Microbiology Jul 2001Mycoplasma arginini, M. fermentans, M. hyorhinis, M. orale, and Acholeplasma laidlawii are the members of the class Mollicutes most commonly found in contaminated cell...
Mycoplasma arginini, M. fermentans, M. hyorhinis, M. orale, and Acholeplasma laidlawii are the members of the class Mollicutes most commonly found in contaminated cell cultures. Previous studies have shown that the published PCR primer pairs designed to detect mollicutes in cell cultures are not entirely specific. The 16S rRNA gene, the 16S-23S rRNA intergenic spacer region, and the 5' end of the 23S rRNA gene, as a whole, are promising targets for design of mollicute species-specific primer pairs. We analyzed the 16S rRNA genes, the 16S-23S rRNA intergenic spacer regions, and the 5' end of the 23S rRNA genes of these mollicutes and developed PCR methods for species identification based on these regions. Using high melting temperatures, we developed a rapid-cycle PCR for detection and identification of contaminant mollicutes. Previously published, putative mollicute-specific primers amplified DNA from 73 contaminated cell lines, but the presence of mollicutes was confirmed by species-specific PCR in only 60. Sequences of the remaining 13 amplicons were identified as those of gram-positive bacterial species. Species-specific PCR primers are needed to confirm the presence of mollicutes in specimens and for identification, if required.
Topics: Cell Line; DNA Primers; DNA, Ribosomal Spacer; Humans; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Ribosomal, 16S; RNA, Ribosomal, 23S; Sensitivity and Specificity; Sequence Analysis, DNA; Species Specificity; Tenericutes
PubMed: 11425741
DOI: 10.1128/AEM.67.7.3195-3200.2001 -
Journal of Bacteriology Jul 1981Acetate kinase activity was assayed in 13 mycoplasmas. Nine species exhibited the enzymic activity in the direction of either synthesis of acetylphosphate or adenosine... (Comparative Study)
Comparative Study
Acetate kinase activity was assayed in 13 mycoplasmas. Nine species exhibited the enzymic activity in the direction of either synthesis of acetylphosphate or adenosine triphosphate. On the other hand Mycoplasma orale, Mycoplasma arthritidis, Ureaplasma urealyticum (10 serotypes), and two strains of Anaeroplasma species exhibited only minimal levels of the enzymic activity. In these four species, the enzyme does not seem to play a key role in adenosine triphosphate formation.
Topics: Acetate Kinase; Acholeplasma; Adenosine Triphosphate; Mycoplasma; Phosphotransferases; Species Specificity; Ureaplasma
PubMed: 6263869
DOI: 10.1128/jb.147.1.271-273.1981 -
Zhongguo Yao Li Xue Bao = Acta... Nov 1999To determine the susceptibilities of Mycoplasma and Ureaplasma to (S)-(-)-9-fluoro-2,3-dihydro-3-methyl-10... (Comparative Study)
Comparative Study
Antimycoplasmal activities of (S)-(-)-9-fluoro-2,3-dihydro-3-methyl-10 -[4-(2-pyridyl)-1-piperazinyl]-7-oxo-7H-pyrido[1,2,3-de][1,4]benzoxazine -6-carboxylic acid (YH-6) in comparison with other antibiotics in vitro.
AIM
To determine the susceptibilities of Mycoplasma and Ureaplasma to (S)-(-)-9-fluoro-2,3-dihydro-3-methyl-10 -[4-(2-pyridyl)-1-piperazinyl]-7-oxo-7H-pyrido[1,2,3-de][1,4]benzoxazine -6-carboxylic acid (YH-6) and to compare it with those referential quinolones, macrolides, and tetracyclines.
METHODS
The minimum inhibitory concentration (MIC) were determined by microdilution method in vitro.
RESULTS
The MIC of YH-6 for Ureaplasma urealyticum (Uu: 250 micrograms.L-1), Mycoplasma hominis (Mh: 500 micrograms.L-1), M orale (Mo: 125 micrograms.L-1) and M salivarium (Ms: 125 micrograms.L-1) were closely similar to those of macrolides (erythromycin and leucomycin) and were 2-8 folds greater than those of ofloxacin (Ofl). Uu and Mh easily induced resistance to erythromycin and tetracycline. They did not easily form resistance to quinolone (YH-6, Ofl), josamycin and tylosin. Tetracycline-resistance (Tcr) or erythromycin-resistance (EMr) strains of Uu (or Mh) had cross-resistance to erythromycin or tetracycline. However, they had no cross-resistance to quinolone, josamycin and tylosin.
CONCLUSION
YH-6 was a highly active quinolone against Mycoplasma, but could hardly induce resistance to Uu. EMr- or Tcr- strains of Uu (or Mh) had no cross-resistance to YH-6.
Topics: Anti-Bacterial Agents; Drug Resistance, Microbial; Erythromycin; Microbial Sensitivity Tests; Mycoplasma; Oxazines; Piperazines; Tetracycline; Ureaplasma
PubMed: 11270970
DOI: No ID Found -
Infection and Immunity Jun 1980The ability of a mouse mammary tumor cell line to abrogate antibody neutralization of vesicular stomatitis virus was shown to be due to the presence of mycoplasma. The...
The ability of a mouse mammary tumor cell line to abrogate antibody neutralization of vesicular stomatitis virus was shown to be due to the presence of mycoplasma. The mycoplasma was isolated from the cell line and typed as Mycoplasma orale. Colonies of this mycoplasma were used to deliberately infect cell cultures which then gained the capacity to reactivate antibody-neutralized virus. The extent of the reactivation depended on the source of neutralizing antiserum. Other species of mycoplasma were tested and were found to reactivate neutralized virus, indicating that this may be a general phenomenon of mycoplasma contamination.
Topics: Animals; Antibodies, Viral; Antigen-Antibody Reactions; Cell Line; Mink; Mycoplasma; Neutralization Tests; Vesicular stomatitis Indiana virus; Virus Activation
PubMed: 6249748
DOI: 10.1128/iai.28.3.649-653.1980 -
Journal of Clinical Pathology Nov 1967A controlled cultural and serological investigation of mycoplasmas in recurrent oral ulceration was undertaken. No evidence for an aetiological relationship between oral...
A controlled cultural and serological investigation of mycoplasmas in recurrent oral ulceration was undertaken. No evidence for an aetiological relationship between oral mycoplasmas and this condition was obtained.Mycoplasmas, principally M. orale, were frequently recovered from the oral cavities of individuals with natural teeth, but rarely from edentulous subjects.
Topics: Adolescent; Adult; Aged; Antibodies; Child; Female; Humans; Male; Middle Aged; Mouth Diseases; Mycoplasma; Stomatitis, Aphthous
PubMed: 5614072
DOI: 10.1136/jcp.20.6.865