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Article

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

1
Medical Mission Institute, 97074 Würzburg, Germany
2
Public Health Laboratory, Secretariat of Health, Department Cesar, Valledupar 20000001, Colombia
3
Organization Wiwa Yugumaiun Bunkauanarrua Tayrona (OWYBT), Department Health Advocacy, Valledupar 2000001, Colombia
4
TibMolBiol, 10103 Berlin, Germany
5
Institute for Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, Germany
6
Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: André Talvani
Microorganisms 2022, 10(8), 1554; https://doi.org/10.3390/microorganisms10081554
Received: 25 May 2022 / Revised: 27 July 2022 / Accepted: 29 July 2022 / Published: 31 July 2022
(This article belongs to the Special Issue Recent Advances in Trypanosoma cruzi Infections)
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. View Full-Text
Keywords: Chagas; pre-analytics; evaluation; diagnostic accuracy; test comparison Chagas; pre-analytics; evaluation; diagnostic accuracy; test comparison
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MDPI and ACS Style

Kann, S.; Zabala-Monterroza, W.; García, C.; Concha, G.; Landt, O.; Hahn, A.; Weinreich, F.; Frickmann, H. Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR. Microorganisms 2022, 10, 1554. https://doi.org/10.3390/microorganisms10081554

AMA Style

Kann S, Zabala-Monterroza W, García C, Concha G, Landt O, Hahn A, Weinreich F, Frickmann H. Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR. Microorganisms. 2022; 10(8):1554. https://doi.org/10.3390/microorganisms10081554

Chicago/Turabian Style

Kann, Simone, Wendy Zabala-Monterroza, Cenia García, Gustavo Concha, Olfert Landt, Andreas Hahn, Felix Weinreich, and Hagen Frickmann. 2022. "Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity of Trypanosoma cruzi-Specific Real-Time PCR" Microorganisms 10, no. 8: 1554. https://doi.org/10.3390/microorganisms10081554

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