Detection of Anaplasma phagocytophilum in animals by real‐time polymerase chain reaction
Identifieur interne : 000A14 ( Main/Exploration ); précédent : 000A13; suivant : 000A15Detection of Anaplasma phagocytophilum in animals by real‐time polymerase chain reaction
Auteurs : Dagmar Hulínská [République tchèque] ; Kate Ina Langrová [République tchèque] ; Milan Pej Och [République tchèque] ; Ivan Pavlásek [République tchèque]Source :
- APMIS [ 0903-4641 ] ; 2004-04.
English descriptors
Abstract
The aim of this study was to detect Anaplasma phagocytophilum in wild and domesticated animals and to identify the phylogenetic relationships of different strains of this bacterium. We adapted six published conventional methods targeting 16S fragments for real‐time polymerase chain reaction. Initial screening of samples from 419 animals found 37 Anaplasma positives, later confirmed with several different primers and a TaqMan probe. We also performed DNA quantification and melting curve analysis. The nucleic acid of Anaplasma sp. was detected in a higher percentage of cases in members of the deer family, hares, bank voles and mice (12.5∼15%) than in foxes, boars, cows, and horses (around 4∼6%). We also performed blood analysis of cows, horses, mice, and ticks removed from animals, evaluating the presence of antibodies against granulocytic Anaplasma sp. Finally, we subjected 11 randomly selected PCR amplified products to direct sequencing and we constructed the corresponding phylogenetic tree with respect to the Ehrlichia equi sequence, homologous to the human granulocytic ehrlichiosis agent. Mutual identity of the sequencing ranged from 99% to 100%.
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DOI: 10.1111/j.1600-0463.2004.apm11204-0503.x
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temperature</term><term>real‐time PCR</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The aim of this study was to detect Anaplasma phagocytophilum in wild and domesticated animals and to identify the phylogenetic relationships of different strains of this bacterium. We adapted six published conventional methods targeting 16S fragments for real‐time polymerase chain reaction. Initial screening of samples from 419 animals found 37 Anaplasma positives, later confirmed with several different primers and a TaqMan probe. We also performed DNA quantification and melting curve analysis. The nucleic acid of Anaplasma sp. was detected in a higher percentage of cases in members of the deer family, hares, bank voles and mice (12.5∼15%) than in foxes, boars, cows, and horses (around 4∼6%). We also performed blood analysis of cows, horses, mice, and ticks removed from animals, evaluating the presence of antibodies against granulocytic Anaplasma sp. Finally, we subjected 11 randomly selected PCR amplified products to direct sequencing and we constructed the corresponding phylogenetic tree with respect to the Ehrlichia equi sequence, homologous to the human granulocytic ehrlichiosis agent. Mutual identity of the sequencing ranged from 99% to 100%.</div></front></TEI><affiliations><list><country><li>République tchèque</li></country><region><li>Bohême centrale</li></region><settlement><li>Prague</li></settlement></list><tree><country name="République tchèque"><region name="Bohême centrale"><name sortKey="Hulinska, Dagmar" sort="Hulinska, Dagmar" uniqKey="Hulinska D" first="Dagmar" last="Hulínská">Dagmar Hulínská</name></region><name sortKey="Hulinska, Dagmar" sort="Hulinska, Dagmar" uniqKey="Hulinska D" first="Dagmar" last="Hulínská">Dagmar Hulínská</name><name sortKey="Langrova, Kate Ina" sort="Langrova, Kate Ina" uniqKey="Langrova K" first="Kate Ina" last="Langrová">Kate Ina Langrová</name><name sortKey="Pavlasek, Ivan" sort="Pavlasek, Ivan" uniqKey="Pavlasek I" first="Ivan" last="Pavlásek">Ivan Pavlásek</name><name sortKey="Pej Och, Milan" sort="Pej Och, Milan" uniqKey="Pej Och M" first="Milan" last="Pej Och">Milan Pej Och</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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