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Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR

For the molecular diagnosis of Chagas disease by real-time PCR (polymerase chain reaction), optimization of diagnostic accuracy is desirable. The detection limit of real-time PCR assays for the diagnosis of Trypanosoma cruzi in human serum is affected by various influences including the choice of th...

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Autores principales: Kann, Simone, Zabala-Monterroza, Wendy, García, Cenia, Concha, Gustavo, Landt, Olfert, Hahn, Andreas, Weinreich, Felix, Frickmann, Hagen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9414588/
https://www.ncbi.nlm.nih.gov/pubmed/36013972
http://dx.doi.org/10.3390/microorganisms10081554
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author Kann, Simone
Zabala-Monterroza, Wendy
García, Cenia
Concha, Gustavo
Landt, Olfert
Hahn, Andreas
Weinreich, Felix
Frickmann, Hagen
author_facet Kann, Simone
Zabala-Monterroza, Wendy
García, Cenia
Concha, Gustavo
Landt, Olfert
Hahn, Andreas
Weinreich, Felix
Frickmann, Hagen
author_sort Kann, Simone
collection PubMed
description For the molecular diagnosis of Chagas disease by real-time PCR (polymerase chain reaction), optimization of diagnostic accuracy is desirable. The detection limit of real-time PCR assays for the diagnosis of Trypanosoma cruzi in human serum is affected by various influences including the choice of the nucleic acid extraction assay. In this study, three nucleic acid extraction assays were compared regarding their influence on the sensitivity of a T. cruzi-specific real-time PCR with 62 reference sera containing T. cruzi target DNA (deoxyribonucleotide acid). More than 95% of the positive sera were correctly identified after all three nucleic acid extraction strategies with a detection rate ranging from 96.8% (60/62) for the worst assay to 100% (62/62) for the best one. A matched pairs analysis for the comparison of the cycle threshold (Ct) values obtained with the 59 reference samples with positive real-time PCR results after all three nucleic acid extraction schemes indicated differences in a range of about 3 Ct steps. Summarized, all three compared nucleic acid extraction schemes were basically suitable for T. cruzi-specific PCR from serum with some minor differences. However, in the case of low quantities of circulating parasite DNA in the serum of a patient with Chagas disease, even minor effects can make a difference in the individual diagnosis.
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spelling pubmed-94145882022-08-27 Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR Kann, Simone Zabala-Monterroza, Wendy García, Cenia Concha, Gustavo Landt, Olfert Hahn, Andreas Weinreich, Felix Frickmann, Hagen Microorganisms Article For the molecular diagnosis of Chagas disease by real-time PCR (polymerase chain reaction), optimization of diagnostic accuracy is desirable. The detection limit of real-time PCR assays for the diagnosis of Trypanosoma cruzi in human serum is affected by various influences including the choice of the nucleic acid extraction assay. In this study, three nucleic acid extraction assays were compared regarding their influence on the sensitivity of a T. cruzi-specific real-time PCR with 62 reference sera containing T. cruzi target DNA (deoxyribonucleotide acid). More than 95% of the positive sera were correctly identified after all three nucleic acid extraction strategies with a detection rate ranging from 96.8% (60/62) for the worst assay to 100% (62/62) for the best one. A matched pairs analysis for the comparison of the cycle threshold (Ct) values obtained with the 59 reference samples with positive real-time PCR results after all three nucleic acid extraction schemes indicated differences in a range of about 3 Ct steps. Summarized, all three compared nucleic acid extraction schemes were basically suitable for T. cruzi-specific PCR from serum with some minor differences. However, in the case of low quantities of circulating parasite DNA in the serum of a patient with Chagas disease, even minor effects can make a difference in the individual diagnosis. MDPI 2022-07-31 /pmc/articles/PMC9414588/ /pubmed/36013972 http://dx.doi.org/10.3390/microorganisms10081554 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kann, Simone
Zabala-Monterroza, Wendy
García, Cenia
Concha, Gustavo
Landt, Olfert
Hahn, Andreas
Weinreich, Felix
Frickmann, Hagen
Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title_full Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title_fullStr Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title_full_unstemmed Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title_short Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR
title_sort comparison of the influence of different nucleic acid extraction assays on the sensitivity of trypanosoma cruzi-specific real-time pcr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9414588/
https://www.ncbi.nlm.nih.gov/pubmed/36013972
http://dx.doi.org/10.3390/microorganisms10081554
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