ArticlePloS one2018
Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 34 citations in OpenAlex.
- Consensus framework for developing a target product profile of real-time PCR in Chagas disease therapeutic monitoring.PLoS neglected tropical diseases · 2026Article
- Chagas Disease in a Non-Endemic Setting: Clinical Profile, Treatment Outcomes, and Predictors of Cure in a 15-Year Cohort Study.Tropical medicine and infectious disease · 2025Article
- Parasite Burden ofMicroorganisms · 2024Article
- Article
- Validation of the NAT Chagas IVD Kit for the Detection and Quantification ofLife (Basel, Switzerland) · 2023Article
- Comparative Assessment of Two Commercial Real-Time PCR Assays for the Diagnosis ofMicroorganisms · 2023Article
- Effects of Acute and Chronic Trypanosoma cruzi Infection on Pregnancy Outcomes in Mice: Parasite Transmission, Mortality, Delayed Growth, and Organ Damage in Pups.The American journal of pathology · 2023Article
- Molecular Detection of Neglected Tropical Diseases: The Case for Automated Near-Point-of-Care Diagnosis of Leishmaniasis.The American journal of tropical medicine and hygiene · 2023Article
- Parasitemia Levels in Trypanosoma cruzi Infection in Spain, an Area Where the Disease Is Not Endemic: Trends by Different Molecular Approaches.Microbiology spectrum · 2022Article
- Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity ofMicroorganisms · 2022Article
- Worldwide Control and Management of Chagas Disease in a New Era of Globalization: a Close Look at Congenital Trypanosoma cruzi Infection.Clinical microbiology reviews · 2022Review
- Parasitological, serological and molecular diagnosis of acute and chronic Chagas disease: from field to laboratory.Memorias do Instituto Oswaldo Cruz · 2022Article
- Combined use of real-time PCR and serological techniques for improved surveillance of chronic and acute American trypanosomiasis in dogs and their owners from an endemic rural area of Neotropical Mexico.Current research in parasitology & vector-borne diseases · 2022Article
- Prospective multicenter evaluation of real time PCR Kit prototype for early diagnosis of congenital Chagas disease.EBioMedicine · 2021Article
- Evaluation of the Performance of the Loopamp Trypanosoma cruzi Detection Kit for the Diagnosis of Chagas Disease in an Area Where It Is Not Endemic, Spain.Journal of clinical microbiology · 2021Article
- Performance Evaluation of a Commercial Real-Time PCR Assay and of an In-House Real-Time PCR forMicroorganisms · 2020Article
- Trypanosoma cruzi loop-mediated isothermal amplification (Trypanosoma cruzi Loopamp) kit for detection of congenital, acute and Chagas disease reactivation.PLoS neglected tropical diseases · 2020Article
- Plasma-Derived Extracellular Vesicles as Potential Biomarkers in Heart Transplant Patient with Chronic Chagas Disease.Emerging infectious diseases · 2020Article
- Congenital Chagas disease: Updated recommendations for prevention, diagnosis, treatment, and follow-up of newborns and siblings, girls, women of childbearing age, and pregnant women.PLoS neglected tropical diseases · 2019Article
- The burden of congenital Chagas disease and implementation of molecular diagnostic tools in Latin America.BMJ global health · 2018Article
Corrections and comments
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Authors and funding
13 authors at 5 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPolymerase chain reaction (PCR) has become a useful tool for the diagnosis of Trypanosoma cruzi infection. The development of automated DNA extraction methodologies and PCR systems is an important step toward the standardization of protocols in routine diagnosis. To date, there are only two commercially available Real-Time PCR assays for the routine laboratory detection of T. cruzi DNA in clinical samples: TCRUZIDNA.CE (Diagnostic Bioprobes Srl) and RealCycler CHAG (Progenie Molecular). Our aim was to evaluate the RealCycler CHAG assay taking into account the whole process. METHODOLOGY/PRINCIPAL
findingsWe assessed the usefulness of an automated DNA extraction system based on magnetic particles (EZ1 Virus Mini Kit v2.0, Qiagen) combined with a commercially available Real-Time PCR assay targeting satellite DNA (SatDNA) of T. cruzi (RealCycler CHAG), a methodology used for routine diagnosis in our hospital. It was compared with a well-known strategy combining a commercial DNA isolation kit based on silica columns (High Pure PCR Template Preparation Kit, Roche Diagnostics) with an in-house Real-Time PCR targeting SatDNA. The results of the two methodologies were in almost perfect agreement, indicating they can be used interchangeably. However, when variations in protocol factors were applied (sample treatment, extraction method and Real-Time PCR), the results were less convincing. A comprehensive fine-tuning of the whole procedure is the key to successful results. Guanidine EDTA-blood (GEB) samples are not suitable for DNA extraction based on magnetic particles due to inhibition, at least when samples are not processed immediately. CONCLUSIONS/SIGNIFICANCE: This is the first study to evaluate the RealCycler CHAG assay taking into account the overall process, including three variables (sample treatment, extraction method and Real-Time PCR). Our findings may contribute to the harmonization of protocols between laboratories and to a wider application of Real-Time PCR in molecular diagnostic laboratories associated with health centers.
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