A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application

BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are ava...
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Autor*in:	Aldo Ummarino [verfasserIn] Michele Caputo [verfasserIn] Francesco Antonio Tucci [verfasserIn] Gaetano Pezzicoli [verfasserIn] Ada Piepoli [verfasserIn] Annamaria Gentile [verfasserIn] Tiziana Latiano [verfasserIn] Anna Panza [verfasserIn] Nicholas Calà [verfasserIn] Antonio Pio Ceglia [verfasserIn] Giovanni Pistoio [verfasserIn] Vincenzo Troiano [verfasserIn] Michela Pucatti [verfasserIn] Anna Latiano [verfasserIn] Angelo Andriulli [verfasserIn] Antonio Tucci [verfasserIn] Orazio Palmieri [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2022

Schlagwörter:	Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA

Übergeordnetes Werk:	In: Frontiers in Microbiology - Frontiers Media S.A., 2011, 13(2022)
Übergeordnetes Werk:	volume:13 ; year:2022

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fmicb.2022.1028988

Katalog-ID:	DOAJ083827846

Internformat


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520			\|a BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application.
650		4	\|a Enterobiasis vermicularis
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700	0		\|a Francesco Antonio Tucci \|e verfasserin \|4 aut
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allfields	10.3389/fmicb.2022.1028988 doi (DE-627)DOAJ083827846 (DE-599)DOAJac21c740ed344e63bbebf5c82e32598f DE-627 ger DE-627 rakwb eng QR1-502 Aldo Ummarino verfasserin aut A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application. Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology Michele Caputo verfasserin aut Francesco Antonio Tucci verfasserin aut Gaetano Pezzicoli verfasserin aut Ada Piepoli verfasserin aut Annamaria Gentile verfasserin aut Tiziana Latiano verfasserin aut Anna Panza verfasserin aut Nicholas Calà verfasserin aut Antonio Pio Ceglia verfasserin aut Giovanni Pistoio verfasserin aut Vincenzo Troiano verfasserin aut Michela Pucatti verfasserin aut Anna Latiano verfasserin aut Angelo Andriulli verfasserin aut Antonio Tucci verfasserin aut Orazio Palmieri verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 13(2022) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:13 year:2022 https://doi.org/10.3389/fmicb.2022.1028988 kostenfrei https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f kostenfrei https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2022
spelling	10.3389/fmicb.2022.1028988 doi (DE-627)DOAJ083827846 (DE-599)DOAJac21c740ed344e63bbebf5c82e32598f DE-627 ger DE-627 rakwb eng QR1-502 Aldo Ummarino verfasserin aut A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application. Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology Michele Caputo verfasserin aut Francesco Antonio Tucci verfasserin aut Gaetano Pezzicoli verfasserin aut Ada Piepoli verfasserin aut Annamaria Gentile verfasserin aut Tiziana Latiano verfasserin aut Anna Panza verfasserin aut Nicholas Calà verfasserin aut Antonio Pio Ceglia verfasserin aut Giovanni Pistoio verfasserin aut Vincenzo Troiano verfasserin aut Michela Pucatti verfasserin aut Anna Latiano verfasserin aut Angelo Andriulli verfasserin aut Antonio Tucci verfasserin aut Orazio Palmieri verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 13(2022) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:13 year:2022 https://doi.org/10.3389/fmicb.2022.1028988 kostenfrei https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f kostenfrei https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2022
allfields_unstemmed	10.3389/fmicb.2022.1028988 doi (DE-627)DOAJ083827846 (DE-599)DOAJac21c740ed344e63bbebf5c82e32598f DE-627 ger DE-627 rakwb eng QR1-502 Aldo Ummarino verfasserin aut A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application. Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology Michele Caputo verfasserin aut Francesco Antonio Tucci verfasserin aut Gaetano Pezzicoli verfasserin aut Ada Piepoli verfasserin aut Annamaria Gentile verfasserin aut Tiziana Latiano verfasserin aut Anna Panza verfasserin aut Nicholas Calà verfasserin aut Antonio Pio Ceglia verfasserin aut Giovanni Pistoio verfasserin aut Vincenzo Troiano verfasserin aut Michela Pucatti verfasserin aut Anna Latiano verfasserin aut Angelo Andriulli verfasserin aut Antonio Tucci verfasserin aut Orazio Palmieri verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 13(2022) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:13 year:2022 https://doi.org/10.3389/fmicb.2022.1028988 kostenfrei https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f kostenfrei https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2022
allfieldsGer	10.3389/fmicb.2022.1028988 doi (DE-627)DOAJ083827846 (DE-599)DOAJac21c740ed344e63bbebf5c82e32598f DE-627 ger DE-627 rakwb eng QR1-502 Aldo Ummarino verfasserin aut A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application. Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology Michele Caputo verfasserin aut Francesco Antonio Tucci verfasserin aut Gaetano Pezzicoli verfasserin aut Ada Piepoli verfasserin aut Annamaria Gentile verfasserin aut Tiziana Latiano verfasserin aut Anna Panza verfasserin aut Nicholas Calà verfasserin aut Antonio Pio Ceglia verfasserin aut Giovanni Pistoio verfasserin aut Vincenzo Troiano verfasserin aut Michela Pucatti verfasserin aut Anna Latiano verfasserin aut Angelo Andriulli verfasserin aut Antonio Tucci verfasserin aut Orazio Palmieri verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 13(2022) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:13 year:2022 https://doi.org/10.3389/fmicb.2022.1028988 kostenfrei https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f kostenfrei https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2022
allfieldsSound	10.3389/fmicb.2022.1028988 doi (DE-627)DOAJ083827846 (DE-599)DOAJac21c740ed344e63bbebf5c82e32598f DE-627 ger DE-627 rakwb eng QR1-502 Aldo Ummarino verfasserin aut A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application. Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology Michele Caputo verfasserin aut Francesco Antonio Tucci verfasserin aut Gaetano Pezzicoli verfasserin aut Ada Piepoli verfasserin aut Annamaria Gentile verfasserin aut Tiziana Latiano verfasserin aut Anna Panza verfasserin aut Nicholas Calà verfasserin aut Antonio Pio Ceglia verfasserin aut Giovanni Pistoio verfasserin aut Vincenzo Troiano verfasserin aut Michela Pucatti verfasserin aut Anna Latiano verfasserin aut Angelo Andriulli verfasserin aut Antonio Tucci verfasserin aut Orazio Palmieri verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 13(2022) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:13 year:2022 https://doi.org/10.3389/fmicb.2022.1028988 kostenfrei https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f kostenfrei https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 13 2022
language	English
source	In Frontiers in Microbiology 13(2022) volume:13 year:2022
sourceStr	In Frontiers in Microbiology 13(2022) volume:13 year:2022
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA Microbiology
isfreeaccess_bool	true
container_title	Frontiers in Microbiology
authorswithroles_txt_mv	Aldo Ummarino @@aut@@ Michele Caputo @@aut@@ Francesco Antonio Tucci @@aut@@ Gaetano Pezzicoli @@aut@@ Ada Piepoli @@aut@@ Annamaria Gentile @@aut@@ Tiziana Latiano @@aut@@ Anna Panza @@aut@@ Nicholas Calà @@aut@@ Antonio Pio Ceglia @@aut@@ Giovanni Pistoio @@aut@@ Vincenzo Troiano @@aut@@ Michela Pucatti @@aut@@ Anna Latiano @@aut@@ Angelo Andriulli @@aut@@ Antonio Tucci @@aut@@ Orazio Palmieri @@aut@@
publishDateDaySort_date	2022-01-01T00:00:00Z
hierarchy_top_id	642889384
id	DOAJ083827846
language_de	englisch
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Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. 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callnumber-first	Q - Science
author	Aldo Ummarino
spellingShingle	Aldo Ummarino misc QR1-502 misc Enterobiasis vermicularis misc PCR misc stool (DNA) test misc pinworm infection misc ribosomal DNA–rDNA misc Microbiology A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application
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topic_title	QR1-502 A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application Enterobiasis vermicularis PCR stool (DNA) test pinworm infection ribosomal DNA–rDNA
topic	misc QR1-502 misc Enterobiasis vermicularis misc PCR misc stool (DNA) test misc pinworm infection misc ribosomal DNA–rDNA misc Microbiology
topic_unstemmed	misc QR1-502 misc Enterobiasis vermicularis misc PCR misc stool (DNA) test misc pinworm infection misc ribosomal DNA–rDNA misc Microbiology
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title	A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application
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title_full	A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application
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author_browse	Aldo Ummarino Michele Caputo Francesco Antonio Tucci Gaetano Pezzicoli Ada Piepoli Annamaria Gentile Tiziana Latiano Anna Panza Nicholas Calà Antonio Pio Ceglia Giovanni Pistoio Vincenzo Troiano Michela Pucatti Anna Latiano Angelo Andriulli Antonio Tucci Orazio Palmieri
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doi_str_mv	10.3389/fmicb.2022.1028988
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title_sort	pcr-based method for the diagnosis of enterobius vermicularis in stool samples, specifically designed for clinical application
callnumber	QR1-502
title_auth	A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application
abstract	BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application.
abstractGer	BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application.
abstract_unstemmed	BackgroundEnterobius vermicularis (E. vermicularis) is a nematode that infects up to 200 million people worldwide, despite effective medications being available. Conventional diagnostic tests are hindered by low sensitivity and poor patient compliance. Furthermore, no biomolecular techniques are available for clinical application. The aim of this study was to develop a procedure specifically designed for clinical application to detect E. vermicularis by means of PCR.Materials and methodsTwo subject groups were taken into account: a group of 27 infected patients and a control group of 27 healthy subjects. A nested-PCR was performed on fecal samples to detect E. vermicularis. Due to the intrinsic difficulties of the fecal matrix, several countermeasures were adopted to ensure the efficient performance of the method: (a) a large amount of feces for the extraction process (20 g instead of 200 mg); (b) a combination of chemical and physical treatments to grind the fecal matrix; (c) an additional purification process for the negative samples after the first nested-PCR; and (d) the selection of a very specific target region for the PCR.ResultsDue to the lack of overlap with other organisms, a sequence of the 5S ribosomal DNA (rDNA) spacer region including the tract SL1 was chosen to design appropriate external and internal primers. The first nested-PCR detected E.vermicularis in 19/27 samples from infected patients. After further purification, 5/8 of the negative samples resulted positive at the second PCR. Conversely, all the samples from healthy controls resulted negative to both PCRs. Sensitivity and specificity of the method were, respectively, 88.9% and 100%.ConclusionThe results prove the high diagnostic accuracy of the proposed method, addressing and overcoming the challenges posed by both conventional tests and PCR-based approaches. Therefore, the method can be proposed for clinical application.
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title_short	A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application
url	https://doi.org/10.3389/fmicb.2022.1028988 https://doaj.org/article/ac21c740ed344e63bbebf5c82e32598f https://www.frontiersin.org/articles/10.3389/fmicb.2022.1028988/full https://doaj.org/toc/1664-302X
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author2	Michele Caputo Francesco Antonio Tucci Gaetano Pezzicoli Ada Piepoli Annamaria Gentile Tiziana Latiano Anna Panza Nicholas Calà Antonio Pio Ceglia Giovanni Pistoio Vincenzo Troiano Michela Pucatti Anna Latiano Angelo Andriulli Antonio Tucci Orazio Palmieri
author2Str	Michele Caputo Francesco Antonio Tucci Gaetano Pezzicoli Ada Piepoli Annamaria Gentile Tiziana Latiano Anna Panza Nicholas Calà Antonio Pio Ceglia Giovanni Pistoio Vincenzo Troiano Michela Pucatti Anna Latiano Angelo Andriulli Antonio Tucci Orazio Palmieri
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up_date	2024-07-03T19:39:25.588Z
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A PCR-based method for the diagnosis of Enterobius vermicularis in stool samples, specifically designed for clinical application

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