Broad-range PCR, cloning and sequencing of the full 16S rRNA gene for detection of bacterial DNA in synovial fluid samples of Tunisian patients with reactive and undifferentiated arthritis
Identifieur interne : 000084 ( Main/Exploration ); précédent : 000083; suivant : 000085Broad-range PCR, cloning and sequencing of the full 16S rRNA gene for detection of bacterial DNA in synovial fluid samples of Tunisian patients with reactive and undifferentiated arthritis
Auteurs : Mariam Siala [Tunisie] ; Radhouane Gdoura [Tunisie] ; Hela Fourati [Tunisie] ; Markus Rihl ; Benoit Jaulhac [France] ; Mohamed Younes [Tunisie] ; Jean Sibilia [France] ; Sofien Baklouti [Tunisie] ; Naceur Bargaoui [Tunisie] ; Slaheddine Sellami [Tunisie] ; Abdelghani Sghir [France] ; Adnane Hammami [Tunisie]Source :
- Arthritis Research & Therapy [ 1478-6354 ] ; 2009.
Descripteurs français
- KwdFr :
- ADN bactérien (génétique), ADN bactérien (isolement et purification), ARN ribosomique 16S (génétique), Adulte (MeSH), Adulte d'âge moyen (MeSH), Arthrite (microbiologie), Clonage moléculaire (méthodes), Femelle (MeSH), Humains (MeSH), Mâle (MeSH), Réaction de polymérisation en chaîne (méthodes), Sujet âgé (MeSH), Synovie (microbiologie), Tunisie (MeSH).
- MESH :
- génétique : ADN bactérien, ARN ribosomique 16S.
- isolement et purification : ADN bactérien.
- microbiologie : Arthrite, Synovie.
- méthodes : Clonage moléculaire, Réaction de polymérisation en chaîne.
- Adulte, Adulte d'âge moyen, Femelle, Humains, Mâle, Sujet âgé, Tunisie.
- Wicri :
- geographic : Tunisie.
English descriptors
- KwdEn :
- Adult (MeSH), Aged (MeSH), Arthritis (microbiology), Cloning, Molecular (methods), DNA, Bacterial (genetics), DNA, Bacterial (isolation & purification), Female (MeSH), Humans (MeSH), Male (MeSH), Middle Aged (MeSH), Polymerase Chain Reaction (methods), RNA, Ribosomal, 16S (genetics), Synovial Fluid (microbiology), Tunisia (MeSH).
- MESH :
- chemical , genetics : DNA, Bacterial, RNA, Ribosomal, 16S.
- chemical , isolation & purification : DNA, Bacterial.
- geographic : Tunisia.
- methods : Cloning, Molecular, Polymerase Chain Reaction.
- microbiology : Arthritis, Synovial Fluid.
- Adult, Aged, Female, Humans, Male, Middle Aged.
Abstract
Broad-range rDNA PCR provides an alternative, cultivation-independent approach for identifying bacterial DNA in reactive and other form of arthritis. The aim of this study was to use broad-range rDNA PCR targeting the 16S rRNA gene in patients with reactive and other forms of arthritis and to screen for the presence of DNA from any given bacterial species in synovial fluid (SF) samples.
We examined the SF samples from a total of 27 patients consisting of patients with reactive arthritis (ReA) (n = 5), undifferentiated arthritis (UA) (n = 9), rheumatoid arthritis (n = 7), and osteoarthritis (n = 6) of which the latter two were used as controls. Using broad-range bacterial PCR amplifying a 1400 bp fragment from the 16S rRNA gene, we identified and sequenced at least 24 clones from each SF sample. To identify the corresponding bacteria, DNA sequences were compared to the EMBL (European Molecular Biology Laboratory) database.
Bacterial DNA was identified in 20 of the 27 SF samples (74, 10%). Analysis of a large number of sequences revealed the presence of DNA from more than one single bacterial species in the SF of all patients studied. The nearly complete sequences of the 1400 bp were obtained for most of the detected species. DNA of bacterial species including
Broad-range PCR followed by cloning and sequencing the entire 16S rDNA, allowed the identification of the bacterial DNA environment in the SF samples of arthritic patients. We found a wide spectrum of bacteria including those known to be involved in ReA and others not previously associated with arthritis.
Url:
DOI: 10.1186/ar2748
PubMed: 19570210
PubMed Central: 2745777
Affiliations:
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Le document en format XML
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<author><name sortKey="Rihl, Markus" sort="Rihl, Markus" uniqKey="Rihl M" first="Markus" last="Rihl">Markus Rihl</name>
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<wicri:noCountry code="subfield">30625 Hannover; Germany</wicri:noCountry>
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<author><name sortKey="Jaulhac, Benoit" sort="Jaulhac, Benoit" uniqKey="Jaulhac B" first="Benoit" last="Jaulhac">Benoit Jaulhac</name>
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<country xml:lang="fr">France</country>
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<author><name sortKey="Younes, Mohamed" sort="Younes, Mohamed" uniqKey="Younes M" first="Mohamed" last="Younes">Mohamed Younes</name>
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<country xml:lang="fr">Tunisie</country>
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<author><name sortKey="Sibilia, Jean" sort="Sibilia, Jean" uniqKey="Sibilia J" first="Jean" last="Sibilia">Jean Sibilia</name>
<affiliation wicri:level="1"><nlm:aff id="I4">Laboratoire de Physiopathologie des Interactions Hôte-bactérie, UPRES-EA 3432, Faculté de Médecine, Université Louis-Pasteur, rue Koeberlé, 67000 Strasbourg, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratoire de Physiopathologie des Interactions Hôte-bactérie, UPRES-EA 3432, Faculté de Médecine, Université Louis-Pasteur, rue Koeberlé, 67000 Strasbourg</wicri:regionArea>
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<author><name sortKey="Baklouti, Sofien" sort="Baklouti, Sofien" uniqKey="Baklouti S" first="Sofien" last="Baklouti">Sofien Baklouti</name>
<affiliation wicri:level="1"><nlm:aff id="I2">Service de Rhumatologie Hôpital Hedi Chaker, Avenue Majida Boulila, 3029 Sfax, Tunisie</nlm:aff>
<country xml:lang="fr">Tunisie</country>
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<author><name sortKey="Bargaoui, Naceur" sort="Bargaoui, Naceur" uniqKey="Bargaoui N" first="Naceur" last="Bargaoui">Naceur Bargaoui</name>
<affiliation wicri:level="1"><nlm:aff id="I5">Service de Rhumatologie, EPS Fattouma Bourguiba, Rue 1er Juin, 5019 Monastir, Tunisie</nlm:aff>
<country xml:lang="fr">Tunisie</country>
<wicri:regionArea>Service de Rhumatologie, EPS Fattouma Bourguiba, Rue 1er Juin, 5019 Monastir</wicri:regionArea>
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<author><name sortKey="Sellami, Slaheddine" sort="Sellami, Slaheddine" uniqKey="Sellami S" first="Slaheddine" last="Sellami">Slaheddine Sellami</name>
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<country xml:lang="fr">Tunisie</country>
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<author><name sortKey="Sghir, Abdelghani" sort="Sghir, Abdelghani" uniqKey="Sghir A" first="Abdelghani" last="Sghir">Abdelghani Sghir</name>
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<country xml:lang="fr">France</country>
<wicri:regionArea>CNRS-UMR 8030, CEA-Genoscope, rue Gaston Crémieux, 91000 Évry</wicri:regionArea>
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<affiliation wicri:level="1"><nlm:aff id="I8">University of Evry Val d'Essonne, Boulevard François Mitterrand, 91025 Évry Cedex, 91000 Évry, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>University of Evry Val d'Essonne, Boulevard François Mitterrand, 91025 Évry Cedex, 91000 Évry</wicri:regionArea>
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<country xml:lang="fr">Tunisie</country>
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<term>Aged (MeSH)</term>
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<term>Cloning, Molecular (methods)</term>
<term>DNA, Bacterial (genetics)</term>
<term>DNA, Bacterial (isolation & purification)</term>
<term>Female (MeSH)</term>
<term>Humans (MeSH)</term>
<term>Male (MeSH)</term>
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<term>Arthrite (microbiologie)</term>
<term>Clonage moléculaire (méthodes)</term>
<term>Femelle (MeSH)</term>
<term>Humains (MeSH)</term>
<term>Mâle (MeSH)</term>
<term>Réaction de polymérisation en chaîne (méthodes)</term>
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<term>Tunisie (MeSH)</term>
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<term>Polymerase Chain Reaction</term>
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</profileDesc>
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<front><div type="abstract" xml:lang="en"><sec><title>Introduction</title>
<p>Broad-range rDNA PCR provides an alternative, cultivation-independent approach for identifying bacterial DNA in reactive and other form of arthritis. The aim of this study was to use broad-range rDNA PCR targeting the 16S rRNA gene in patients with reactive and other forms of arthritis and to screen for the presence of DNA from any given bacterial species in synovial fluid (SF) samples.</p>
</sec>
<sec sec-type="methods"><title>Methods</title>
<p>We examined the SF samples from a total of 27 patients consisting of patients with reactive arthritis (ReA) (n = 5), undifferentiated arthritis (UA) (n = 9), rheumatoid arthritis (n = 7), and osteoarthritis (n = 6) of which the latter two were used as controls. Using broad-range bacterial PCR amplifying a 1400 bp fragment from the 16S rRNA gene, we identified and sequenced at least 24 clones from each SF sample. To identify the corresponding bacteria, DNA sequences were compared to the EMBL (European Molecular Biology Laboratory) database.</p>
</sec>
<sec><title>Results</title>
<p>Bacterial DNA was identified in 20 of the 27 SF samples (74, 10%). Analysis of a large number of sequences revealed the presence of DNA from more than one single bacterial species in the SF of all patients studied. The nearly complete sequences of the 1400 bp were obtained for most of the detected species. DNA of bacterial species including <italic>Shigella </italic>
species, <italic>Escherichia </italic>
species, and other coli-form bacteria as well as opportunistic pathogens such as <italic>Stenotrophomonas maltophilia </italic>
and <italic>Achromobacter xylosoxidans </italic>
were shared in all arthritis patients. Among pathogens described to trigger ReA, DNA from <italic>Shigella sonnei </italic>
was found in ReA and UA patients. We also detected DNA from rarely occurring human pathogens such as <italic>Aranicola </italic>
species and <italic>Pantoea ananatis</italic>
. We also found DNA from bacteria so far not described in human infections such as <italic>Bacillus niacini</italic>
, <italic>Paenibacillus humicus</italic>
, <italic>Diaphorobacter </italic>
species and uncultured bacterium genera incertae sedis OP10.</p>
</sec>
<sec><title>Conclusions</title>
<p>Broad-range PCR followed by cloning and sequencing the entire 16S rDNA, allowed the identification of the bacterial DNA environment in the SF samples of arthritic patients. We found a wide spectrum of bacteria including those known to be involved in ReA and others not previously associated with arthritis.</p>
</sec>
</div>
</front>
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</TEI>
<affiliations><list><country><li>France</li>
<li>Tunisie</li>
</country>
</list>
<tree><noCountry><name sortKey="Rihl, Markus" sort="Rihl, Markus" uniqKey="Rihl M" first="Markus" last="Rihl">Markus Rihl</name>
</noCountry>
<country name="Tunisie"><noRegion><name sortKey="Siala, Mariam" sort="Siala, Mariam" uniqKey="Siala M" first="Mariam" last="Siala">Mariam Siala</name>
</noRegion>
<name sortKey="Baklouti, Sofien" sort="Baklouti, Sofien" uniqKey="Baklouti S" first="Sofien" last="Baklouti">Sofien Baklouti</name>
<name sortKey="Bargaoui, Naceur" sort="Bargaoui, Naceur" uniqKey="Bargaoui N" first="Naceur" last="Bargaoui">Naceur Bargaoui</name>
<name sortKey="Fourati, Hela" sort="Fourati, Hela" uniqKey="Fourati H" first="Hela" last="Fourati">Hela Fourati</name>
<name sortKey="Gdoura, Radhouane" sort="Gdoura, Radhouane" uniqKey="Gdoura R" first="Radhouane" last="Gdoura">Radhouane Gdoura</name>
<name sortKey="Hammami, Adnane" sort="Hammami, Adnane" uniqKey="Hammami A" first="Adnane" last="Hammami">Adnane Hammami</name>
<name sortKey="Sellami, Slaheddine" sort="Sellami, Slaheddine" uniqKey="Sellami S" first="Slaheddine" last="Sellami">Slaheddine Sellami</name>
<name sortKey="Younes, Mohamed" sort="Younes, Mohamed" uniqKey="Younes M" first="Mohamed" last="Younes">Mohamed Younes</name>
</country>
<country name="France"><noRegion><name sortKey="Jaulhac, Benoit" sort="Jaulhac, Benoit" uniqKey="Jaulhac B" first="Benoit" last="Jaulhac">Benoit Jaulhac</name>
</noRegion>
<name sortKey="Sghir, Abdelghani" sort="Sghir, Abdelghani" uniqKey="Sghir A" first="Abdelghani" last="Sghir">Abdelghani Sghir</name>
<name sortKey="Sghir, Abdelghani" sort="Sghir, Abdelghani" uniqKey="Sghir A" first="Abdelghani" last="Sghir">Abdelghani Sghir</name>
<name sortKey="Sibilia, Jean" sort="Sibilia, Jean" uniqKey="Sibilia J" first="Jean" last="Sibilia">Jean Sibilia</name>
</country>
</tree>
</affiliations>
</record>
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