Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease
- Autores
- Muñoz Calderon, Arturo Alejandro; Besuschio, Susana Alicia; Wong, Season; Fernández, Marisa; García Casares, Lady Juliette; Giorgio, Patricia; Barcan, Laura A.; Markham, Cole; Liu, Yanwen E.; Alarcón de Noya, Belkisyolé; Longhi, Silvia Andrea; Schijman, Alejandro Gabriel
- Año de publicación
- 2022
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD.
Fil: Muñoz Calderon, Arturo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Besuschio, Susana Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Wong, Season. Ai Biosciences, Inc.; Estados Unidos
Fil: Fernández, Marisa. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; Argentina
Fil: García Casares, Lady Juliette. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Giorgio, Patricia. Hospital Británico de Buenos Aires; Argentina
Fil: Barcan, Laura A.. Hospital Italiano. Departamento de Medicina.; Argentina
Fil: Markham, Cole. Ai Biosciences, Inc.; Estados Unidos
Fil: Liu, Yanwen E.. Ai Biosciences, Inc.; Estados Unidos
Fil: Alarcón de Noya, Belkisyolé. Universidad Central de Venezuela; Venezuela
Fil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina - Materia
-
CHAGAS DISEASE REACTIVATION
CHAGAS-HIV
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
ORALLY TRANSMITTED CHAGAS DISEASE
PRIMARY INFECTION AFTER TRANSPLANT IN SEROPOSITIVE DONOR-SERONEGATIVE RECIPIENTS
REAL-TIME PCR
TRYPANOSOMA CRUZI - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by/2.5/ar/
- Repositorio
.jpg)
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/201412
Ver los metadatos del registro completo
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Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas DiseaseMuñoz Calderon, Arturo AlejandroBesuschio, Susana AliciaWong, SeasonFernández, MarisaGarcía Casares, Lady JulietteGiorgio, PatriciaBarcan, Laura A.Markham, ColeLiu, Yanwen E.Alarcón de Noya, BelkisyoléLonghi, Silvia AndreaSchijman, Alejandro GabrielCHAGAS DISEASE REACTIVATIONCHAGAS-HIVLOOP-MEDIATED ISOTHERMAL AMPLIFICATIONORALLY TRANSMITTED CHAGAS DISEASEPRIMARY INFECTION AFTER TRANSPLANT IN SEROPOSITIVE DONOR-SERONEGATIVE RECIPIENTSREAL-TIME PCRTRYPANOSOMA CRUZIhttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD.Fil: Muñoz Calderon, Arturo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Besuschio, Susana Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Wong, Season. Ai Biosciences, Inc.; Estados UnidosFil: Fernández, Marisa. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; ArgentinaFil: García Casares, Lady Juliette. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Giorgio, Patricia. Hospital Británico de Buenos Aires; ArgentinaFil: Barcan, Laura A.. Hospital Italiano. Departamento de Medicina.; ArgentinaFil: Markham, Cole. Ai Biosciences, Inc.; Estados UnidosFil: Liu, Yanwen E.. Ai Biosciences, Inc.; Estados UnidosFil: Alarcón de Noya, Belkisyolé. Universidad Central de Venezuela; VenezuelaFil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaMultidisciplinary Digital Publishing Institute2022-04-26info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/201412Muñoz Calderon, Arturo Alejandro; Besuschio, Susana Alicia; Wong, Season; Fernández, Marisa; García Casares, Lady Juliette; et al.; Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease; Multidisciplinary Digital Publishing Institute; Microorganisms; 10; 5; 26-4-2022; 1-102076-2607CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.3390/microorganisms10050909info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2076-2607/10/5/909info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:55:58Zoai:ri.conicet.gov.ar:11336/201412instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:55:59.053CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
| dc.title.none.fl_str_mv |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| title |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| spellingShingle |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease Muñoz Calderon, Arturo Alejandro CHAGAS DISEASE REACTIVATION CHAGAS-HIV LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ORALLY TRANSMITTED CHAGAS DISEASE PRIMARY INFECTION AFTER TRANSPLANT IN SEROPOSITIVE DONOR-SERONEGATIVE RECIPIENTS REAL-TIME PCR TRYPANOSOMA CRUZI |
| title_short |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| title_full |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| title_fullStr |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| title_full_unstemmed |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| title_sort |
Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease |
| dc.creator.none.fl_str_mv |
Muñoz Calderon, Arturo Alejandro Besuschio, Susana Alicia Wong, Season Fernández, Marisa García Casares, Lady Juliette Giorgio, Patricia Barcan, Laura A. Markham, Cole Liu, Yanwen E. Alarcón de Noya, Belkisyolé Longhi, Silvia Andrea Schijman, Alejandro Gabriel |
| author |
Muñoz Calderon, Arturo Alejandro |
| author_facet |
Muñoz Calderon, Arturo Alejandro Besuschio, Susana Alicia Wong, Season Fernández, Marisa García Casares, Lady Juliette Giorgio, Patricia Barcan, Laura A. Markham, Cole Liu, Yanwen E. Alarcón de Noya, Belkisyolé Longhi, Silvia Andrea Schijman, Alejandro Gabriel |
| author_role |
author |
| author2 |
Besuschio, Susana Alicia Wong, Season Fernández, Marisa García Casares, Lady Juliette Giorgio, Patricia Barcan, Laura A. Markham, Cole Liu, Yanwen E. Alarcón de Noya, Belkisyolé Longhi, Silvia Andrea Schijman, Alejandro Gabriel |
| author2_role |
author author author author author author author author author author author |
| dc.subject.none.fl_str_mv |
CHAGAS DISEASE REACTIVATION CHAGAS-HIV LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ORALLY TRANSMITTED CHAGAS DISEASE PRIMARY INFECTION AFTER TRANSPLANT IN SEROPOSITIVE DONOR-SERONEGATIVE RECIPIENTS REAL-TIME PCR TRYPANOSOMA CRUZI |
| topic |
CHAGAS DISEASE REACTIVATION CHAGAS-HIV LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ORALLY TRANSMITTED CHAGAS DISEASE PRIMARY INFECTION AFTER TRANSPLANT IN SEROPOSITIVE DONOR-SERONEGATIVE RECIPIENTS REAL-TIME PCR TRYPANOSOMA CRUZI |
| purl_subject.fl_str_mv |
https://purl.org/becyt/ford/3.4 https://purl.org/becyt/ford/3 |
| dc.description.none.fl_txt_mv |
A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD. Fil: Muñoz Calderon, Arturo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Besuschio, Susana Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Wong, Season. Ai Biosciences, Inc.; Estados Unidos Fil: Fernández, Marisa. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; Argentina Fil: García Casares, Lady Juliette. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Giorgio, Patricia. Hospital Británico de Buenos Aires; Argentina Fil: Barcan, Laura A.. Hospital Italiano. Departamento de Medicina.; Argentina Fil: Markham, Cole. Ai Biosciences, Inc.; Estados Unidos Fil: Liu, Yanwen E.. Ai Biosciences, Inc.; Estados Unidos Fil: Alarcón de Noya, Belkisyolé. Universidad Central de Venezuela; Venezuela Fil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina |
| description |
A loop-mediated isothermal amplification assay was evaluated as a surrogate marker of treatment failure in Chagas disease (CD). A convenience series of 18 acute or reactivated CD patients who received anti-parasitic treatment with benznidazole was selected—namely, nine orally infected patients: three people living with HIV and CD reactivation, five chronic CD recipients with reactivation after organ transplantation and one seronegative recipient of a kidney and liver transplant from a CD donor. Fifty-four archival samples (venous blood treated with EDTA or guanidinium hydrochloride-EDTA buffer and cerebrospinal fluid) were extracted using a Spin-column manual kit and tested by T. cruzi Loopamp kit (Tc-LAMP, index test) and standardized real-time PCR (qPCR, comparator test). Of them, 23 samples were also extracted using a novel repurposed 3D printer designed for point-of-care DNA extraction (PrintrLab). The agreement between methods was estimated by Cohen’s kappa index and Bland–Altman plot analysis. The T. cruzi Loopamp kit was as sensitive as qPCR for detecting parasite DNA in samples with parasite loads higher than 0.5 parasite equivalents/mL and infected with different discrete typing units. The agreement between qPCR and Tc-LAMP (Spin-column) or Tc-LAMP (PrintrLab) was excellent, with a mean difference of 0.02 [CI = −0.58–0.62] and −0.04 [CI = −0.45–0.37] and a Cohen’s kappa coefficient of 0.78 [CI = 0.60–0.96] and 0.90 [CI = 0.71 to 1.00], respectively. These findings encourage prospective field studies to validate the use of LAMP as a surrogate marker of treatment failure in CD. |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022-04-26 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/201412 Muñoz Calderon, Arturo Alejandro; Besuschio, Susana Alicia; Wong, Season; Fernández, Marisa; García Casares, Lady Juliette; et al.; Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease; Multidisciplinary Digital Publishing Institute; Microorganisms; 10; 5; 26-4-2022; 1-10 2076-2607 CONICET Digital CONICET |
| url |
http://hdl.handle.net/11336/201412 |
| identifier_str_mv |
Muñoz Calderon, Arturo Alejandro; Besuschio, Susana Alicia; Wong, Season; Fernández, Marisa; García Casares, Lady Juliette; et al.; Loop-Mediated Isothermal Amplification of Trypanosoma cruzi DNA for Point-of-Care Follow-Up of Anti-Parasitic Treatment of Chagas Disease; Multidisciplinary Digital Publishing Institute; Microorganisms; 10; 5; 26-4-2022; 1-10 2076-2607 CONICET Digital CONICET |
| dc.language.none.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/doi/10.3390/microorganisms10050909 info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2076-2607/10/5/909 |
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info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by/2.5/ar/ |
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openAccess |
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https://creativecommons.org/licenses/by/2.5/ar/ |
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application/pdf application/pdf application/pdf application/pdf application/pdf |
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Multidisciplinary Digital Publishing Institute |
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Multidisciplinary Digital Publishing Institute |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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13.070432 |