Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-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 <i>Trypanosoma cruzi</i> in human serum is affected by various influences includi...
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| Format: | Article |
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MDPI AG
2022-07-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/8/1554 |
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| author | Simone Kann Wendy Zabala-Monterroza Cenia García Gustavo Concha Olfert Landt Andreas Hahn Felix Weinreich Hagen Frickmann |
| author_facet | Simone Kann Wendy Zabala-Monterroza Cenia García Gustavo Concha Olfert Landt Andreas Hahn Felix Weinreich Hagen Frickmann |
| author_sort | Simone Kann |
| collection | DOAJ |
| 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 <i>Trypanosoma cruzi</i> 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 <i>T. cruzi</i>-specific real-time PCR with 62 reference sera containing <i>T. cruzi</i> 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 <i>T. cruzi</i>-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. |
| first_indexed | 2024-03-09T12:55:05Z |
| format | Article |
| id | doaj.art-a27363132bd1427d9d447b50f7f3b986 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T12:55:05Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-a27363132bd1427d9d447b50f7f3b9862023-11-30T22:01:39ZengMDPI AGMicroorganisms2076-26072022-07-01108155410.3390/microorganisms10081554Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCRSimone Kann0Wendy Zabala-Monterroza1Cenia García2Gustavo Concha3Olfert Landt4Andreas Hahn5Felix Weinreich6Hagen Frickmann7Medical Mission Institute, 97074 Würzburg, GermanyPublic Health Laboratory, Secretariat of Health, Department Cesar, Valledupar 20000001, ColombiaPublic Health Laboratory, Secretariat of Health, Department Cesar, Valledupar 20000001, ColombiaOrganization Wiwa Yugumaiun Bunkauanarrua Tayrona (OWYBT), Department Health Advocacy, Valledupar 2000001, ColombiaTibMolBiol, 10103 Berlin, GermanyInstitute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, GermanyDepartment of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, GermanyInstitute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, GermanyFor 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 <i>Trypanosoma cruzi</i> 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 <i>T. cruzi</i>-specific real-time PCR with 62 reference sera containing <i>T. cruzi</i> 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 <i>T. cruzi</i>-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.https://www.mdpi.com/2076-2607/10/8/1554Chagaspre-analyticsevaluationdiagnostic accuracytest comparison |
| spellingShingle | Simone Kann Wendy Zabala-Monterroza Cenia García Gustavo Concha Olfert Landt Andreas Hahn Felix Weinreich Hagen Frickmann Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR Microorganisms Chagas pre-analytics evaluation diagnostic accuracy test comparison |
| title | Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR |
| title_full | Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR |
| title_fullStr | Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR |
| title_full_unstemmed | Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR |
| title_short | Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of <i>Trypanosoma cruzi</i>-Specific Real-Time PCR |
| title_sort | comparison of the influence of different nucleic acid extraction assays on the sensitivity of i trypanosoma cruzi i specific real time pcr |
| topic | Chagas pre-analytics evaluation diagnostic accuracy test comparison |
| url | https://www.mdpi.com/2076-2607/10/8/1554 |
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