Comparison of DNA extraction methods and real-time PCR assays for the molecular diagnostics of chronic Chagas disease

Future Microbiol. 2020 Aug:15:1139-1145. doi: 10.2217/fmb-2020-0023.

Abstract

Aim: This work aimed to compare the sensitivity of four protocols for the detection of Trypanosoma cruzi DNA in 98 blood samples from chronic Chagas disease patients. Materials & methods: Two DNA extraction (automated and manual) methods and two T. cruzi satellite DNA qPCRs (with a recent design and the usually used set of primers) were analyzed. Results: Both DNA extraction methods and qPCR assays tested in this work gave comparable qualitative results, although the lowest Ct values were obtained when samples were analyzed using the new set of primers for T. cruzi satellite DNA. Conclusion: Our results encourage the implementation of automated DNA extraction systems and the new T. cruzi qPCR for the molecular diagnostics and treatment response monitoring of chronic Chagas disease patients.

Keywords: Chagas disease; DNA extraction; Trypanosoma cruzi; clinical trials; molecular diagnostics; real-time PCR; treatment monitoring.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chagas Disease / diagnosis*
  • Chagas Disease / parasitology
  • DNA Primers / genetics
  • DNA, Protozoan / genetics
  • DNA, Protozoan / isolation & purification*
  • Genetic Techniques*
  • Humans
  • Pathology, Molecular / instrumentation
  • Pathology, Molecular / methods*
  • Real-Time Polymerase Chain Reaction / instrumentation
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Trypanosoma cruzi / genetics
  • Trypanosoma cruzi / isolation & purification*

Substances

  • DNA Primers
  • DNA, Protozoan