Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach
Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perfo...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Ettore De Canale [verfasserIn] Maria Angela Biasolo [verfasserIn] Andrea Tessari [verfasserIn] Sabrina Bettanello [verfasserIn] Valeria Besutti [verfasserIn] Carlo Mengoli [verfasserIn] |
|---|
| Format: |
E-Artikel |
|---|---|
| Sprache: |
Englisch ; Italienisch |
| Erschienen: |
2009 |
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| Schlagwörter: |
Dientamoeba fragilis, parasitology, fixed stool, direct wet mount, real-time PCR |
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| Übergeordnetes Werk: |
In: Microbiologia Medica - PAGEPress Publications, 2015, 24(2009), 3 |
|---|---|
| Übergeordnetes Werk: |
volume:24 ; year:2009 ; number:3 |
| Links: |
|---|
| DOI / URN: |
10.4081/mm.2009.2524 |
|---|
| Katalog-ID: |
DOAJ065238249 |
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| 520 | |a Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. | ||
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10.4081/mm.2009.2524 doi (DE-627)DOAJ065238249 (DE-599)DOAJa8d780dcf97e493c9bc3f53d974e7a89 DE-627 ger DE-627 rakwb eng ita QR1-502 Ettore De Canale verfasserin aut Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. Dientamoeba fragilis, parasitology, fixed stool, direct wet mount, real-time PCR Microbiology Maria Angela Biasolo verfasserin aut Andrea Tessari verfasserin aut Sabrina Bettanello verfasserin aut Valeria Besutti verfasserin aut Carlo Mengoli verfasserin aut In Microbiologia Medica PAGEPress Publications, 2015 24(2009), 3 (DE-627)1760636096 22806423 nnns volume:24 year:2009 number:3 https://doi.org/10.4081/mm.2009.2524 kostenfrei https://doaj.org/article/a8d780dcf97e493c9bc3f53d974e7a89 kostenfrei http://www.pagepressjournals.org/index.php/mm/article/view/2524 kostenfrei https://doaj.org/toc/2280-6423 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 24 2009 3 |
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10.4081/mm.2009.2524 doi (DE-627)DOAJ065238249 (DE-599)DOAJa8d780dcf97e493c9bc3f53d974e7a89 DE-627 ger DE-627 rakwb eng ita QR1-502 Ettore De Canale verfasserin aut Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. Dientamoeba fragilis, parasitology, fixed stool, direct wet mount, real-time PCR Microbiology Maria Angela Biasolo verfasserin aut Andrea Tessari verfasserin aut Sabrina Bettanello verfasserin aut Valeria Besutti verfasserin aut Carlo Mengoli verfasserin aut In Microbiologia Medica PAGEPress Publications, 2015 24(2009), 3 (DE-627)1760636096 22806423 nnns volume:24 year:2009 number:3 https://doi.org/10.4081/mm.2009.2524 kostenfrei https://doaj.org/article/a8d780dcf97e493c9bc3f53d974e7a89 kostenfrei http://www.pagepressjournals.org/index.php/mm/article/view/2524 kostenfrei https://doaj.org/toc/2280-6423 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ AR 24 2009 3 |
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QR1-502 Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach Dientamoeba fragilis, parasitology, fixed stool, direct wet mount, real-time PCR |
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Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach |
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Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach |
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Ettore De Canale |
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Microbiologia Medica |
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2009 |
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Ettore De Canale Maria Angela Biasolo Andrea Tessari Sabrina Bettanello Valeria Besutti Carlo Mengoli |
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10.4081/mm.2009.2524 |
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real time pcr for dientamoeba fragilis: a comparison between molecular and microscopical approach |
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Real Time PCR for Dientamoeba fragilis: a comparison between molecular and microscopical approach |
| abstract |
Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. |
| abstractGer |
Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. |
| abstract_unstemmed |
Introduction. Dientamoeba fragilis is an intestinal protozoan parasite that is attracting growing interest. Several authors have associated the presence of this faecal agent to numerous intestinal and systemic clinical symptoms, even though asymptomatic carriers have been reported.Actually, to perform the coproparasitology exam for the identification of D. fragilis it is necessary to recur to an expert microscopist. Recently, several real-time TaqMan assays for the detection of D. fragilis has been reported. In this work we validate an original real-time PCR (TaqMan), re-evaluating the traditional microscopic diagnosis. Moreover, the potential for quantitative information, intrinsic of this molecular technique, was preliminarly explored. Methods. Forty-six carriers of D. fragilis were selected from patients referred to the Service of Microbiology of Padova University Hospital. The parasite was preliminarily detected on several occasions in each patient using traditional microscopy. A control group included forty-two healthy volunteers. Real-time PCR was applied to a stool sample from each subject. On the same sample, gel-electrophoresis amplicon detection PCR and microscopic examination were also performed. Results. The sensitivity of real-time PCR was 100%, whereas microscopy showed 93% sensitivity (direct wet mount on fixed stools and trichrome stain), gel-electrophoresis PCR 76% and Giemsa stain 52%.The specificity of all methods was 100%. On the quantitative side, the frequency distribution of the protozoan load of the stools revealed a near to log-normal pattern is apparent for the data yielded by real-time PCR. Conclusions. The real-time PCR was clearly superior to either the gel electrophoresis PCR or the traditional microscopic examination. Just one unpreserved stool sample was adequate to find the parasite. The entire procedure could be performed within a few hours avoiding toxic dyes. |
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Maria Angela Biasolo Andrea Tessari Sabrina Bettanello Valeria Besutti Carlo Mengoli |
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