16S rDNA PCR and denaturing gradient gel electrophoresis; a single generic test for detecting and differentiating Mycoplasma species

McAuliffe, Laura; Ellis, Richard J.; Lawes, Jo R.; Ayling, Roger D.; Nicholas, R. A. J.

doi:10.1099/jmm.0.46058-0

Cited by 148 publications

(142 citation statements)

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“…method (McAuliffe et al, 2005) is a sensitive single generic test capable of detecting and differentiating mycoplasmas to the species level. By DGGE, the ibex isolate was observed to be a Mmc serovar.…”

Section: Discussionmentioning

confidence: 99%

“…Briefly, Mycoplasma DNA was extracted as stated above for the PCR. The PCR and DGGE were performed as described by McAuliffe et al (2005).…”

Section: Polymerase Chain Reaction Denaturing Gradient Gel Electrophomentioning

confidence: 99%

“…Isolation in culture is difficult and requires enriched and selective media, whereas PCR is rapid, sensitive, and specific. The PCR/denaturing gradient gel electrophoresis (DGGE) method offers significant advantages over standard PCR methods because it detects most animal Mycoplasma species including mixed species in a single reaction (McAuliffe et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

Giangaspero

Orusa

Nicholas

et al. 2010

Journal of Wildlife Diseases

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ABSTRACT:In 2005 a Mycoplasma species was isolated from ocular-conjunctival swabs from an adult male Alpine ibex (Capra ibex) from the Valle d'Aosta Region, Northern Italy. The animal suffered from bilateral ocular discharge with diffuse inflammation, severe corneal involvement of the left eye and mild corneal opacity of the right eye. Histologic examination revealed a keratoconjunctivitis characterized by lymphocytic and plasmacellular infiltration. Laboratory investigations of the isolate included culture, transmission electron microscopy, PCR, and denaturing gradient gel electrophoresis, as well as DNA sequencing of the 16S rDNA gene. These tests identified the isolate as Mycoplasma mycoides subsp. capri large-colony serovar, an organism that has occasionally been associated with keratoconjunctivitis in goats. For a correct diagnosis, it is necessary to carry out laboratory investigations, as clinical cases of keratoconjunctivitis in wild ruminants are not always ascribable to Mycoplasma conjunctivae.

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Section: Discussionmentioning

confidence: 99%

“…Briefly, Mycoplasma DNA was extracted as stated above for the PCR. The PCR and DGGE were performed as described by McAuliffe et al (2005).…”

Section: Polymerase Chain Reaction Denaturing Gradient Gel Electrophomentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

Giangaspero

Orusa

Nicholas

et al. 2010

Journal of Wildlife Diseases

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“…The ISR sequences were identical, in accordance with the isolates' high phenotypic similarity. Furthermore, 16S rRNA gene sequence similarity was studied by means of denaturing gradient gel electrophoresis (McAuliffe et al, 2005), which suggested that the isolates' 16S rRNA gene sequences were also identical (data not shown).…”

mentioning

confidence: 99%

Mycoplasma neophronis sp. nov., isolated from the upper respiratory tract of Canarian Egyptian vultures (Neophron percnopterus majorensis)

Suárez-Pérez

Ramírez

Rosales

et al. 2012

International Journal of Systematic and Evolutionary Microbiology

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Six strains with the typical characteristics of mycoplasmas were isolated from the tracheae of six Canarian Egyptian vultures (Neophron percnopterus majorensis). The results of biochemical, serological and molecular genetic studies showed that the isolates were nearly identical and that they could be considered as representing a novel species of the genus Mycoplasma . Colonies possessed the typical fried-egg appearance and electron micrographs revealed a pleomorphic cellular morphology with the lack of a cell wall. The isolates hydrolysed arginine and required sterol for growth but did not ferment glucose or hydrolyse urea. We propose that the isolates be assigned to a novel species,Mycoplasma neophronis sp. nov. The type strain is G.A.T ( = DSM 24097T = ATCC BAA-2157T). The antiserum of strain G.A.T has been deposited in the Mollicutes collection at Purdue University (Indiana, USA).

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“…DGGE is a molecular approach to detect variation between sequences within a population, or even a community. Through this method, the amplified products of the genes of interest will be used in gel electrophoresis under constant heat by increasing concentration of denaturing agents (formamide or urea) (Muyzer et al, 1993;Walter et al, 2000;McAuliffe et al, 2005). Hydrogen bonds between base pairs will be broken by different concentrations of denaturing agents at a constant temperature.…”

Section: Snps Such As Denaturing Gradient Gel Electrophoresis (Dgge) mentioning

confidence: 99%

Molecular characterisation of flowering genes in Pongamia Pinnata

Kamde¹

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Pongamia pinnata (Pongamia) is a multipurpose tree that has been used traditionally for medicine, green manure, insecticide, fuel and also a source of timber. It is an indigenous plant to the Indian subcontinent and Southeast Asia, and has been successfully introduced to Northern Australia, New Zealand and several other countries with humid tropical lowlands. Recently, Pongamia has attracted interest as one of the alternative feedstock for biodiesel productions due to its ability to produce seeds with high oil content. The fatty acid methyl ester (FAME) composition of Pongamia met the biodiesel specification standards of USA (ASTMD 6751), Germany (DIN 51606) and European Standard Organisation (EN 14214), and is considered as one of the most suitable sources of biodiesel. Despite the positive attributes described above, Pongamia is not a domesticated species and further research is required for the selection of elite cultivars with desirable traits optimal for large scale production of Pongamia oil. In addition, limited information is currently available regarding the flowering mechanism of Pongamia at the molecular level that could provide the platform for the development of oil-rich seeds.The aim of this thesis is to provide insight and increase the understanding of the genetic network that leads to flowering in Pongamia. Based on large phenotypic diversity of Pongamia, this information could subsequently be used to determine and select traits with unique flowering time that will allow a harvesting window, where harvesting process will not clash with flowering. The first objective of this project is to identify, isolate and characterise the candidate genes that regulate flowering time in Pongamia. This was achieved using molecular biology techniques and bioinformatic approaches based on the information available on flowering mechanisms of Arabidopsis and soybean in the literature. The second objective of this project is to determine the expression pattern of isolated genes during flowering onset. This was achieved through transcriptome methodologies such as RT-PCR and qRT-PCR. The third objective of this project is to investigate the genetic variety of Pongamia. This was achieved through Single Nucleotide Polymorphism (SNPs) analysis and its correlation with the late flowering phenotype of Pongamia.PpCOL-1 and PpPHYB are the two genes that were investigated in this thesis. The orthologue genes of PpCOL-1 and PpPHYB are involved in the flowering regulation of the annual plant Arabidopsis.Since Arabidopsis and Pongamia are both long day plants, it is postulated that these genes might play a role in the flowering time control in Pongamia. PpCOL-1 and PpPHYB were successfully isolated, characterised and compared to their respective orthologue genes in Arabidopsis, Soybean and Medicago. It was found that the amino acid residues within the domain regions in these genes iii are highly conserved in Pongamia. Interestingly, there were no significant differences in the expression patterns of PpCOL-1 and PpPHYB in...

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16S rDNA PCR and denaturing gradient gel electrophoresis; a single generic test for detecting and differentiating Mycoplasma species

Cited by 148 publications

References 15 publications

Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

Mycoplasma neophronis sp. nov., isolated from the upper respiratory tract of Canarian Egyptian vultures (Neophron percnopterus majorensis)

Molecular characterisation of flowering genes in Pongamia Pinnata

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