ArticlePLoS neglected tropical diseases2017
Analytical sensitivity and specificity of a loop-mediated isothermal amplification (LAMP) kit prototype for detection of Trypanosoma cruzi DNA in human blood samples.
Article in PLoS neglected tropical diseases, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers, 2 of them syntheses that pooled it.
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Who cites it
50 citing papers in PubMed, 2 syntheses or guidelines pooled it, 95 citations in OpenAlex.
- Molecular diagnosis of Chagas disease: a systematic review and meta-analysis.Infectious diseases of poverty · 2023Pooled it
- Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis.International journal of molecular sciences · 2020Pooled it
- A smartphone-based particle diffusometry platform for sub-attomolar detection of Vibrio cholerae in environmental water.Biosensors & bioelectronics · 2020Trial
- Evaluation of point-of-care tests for cutaneous leishmaniasis diagnosis in Kabul, Afghanistan.EBioMedicine · 2018Trial
- Towards point-of-care diagnostics of neglected tropical diseases using nucleic acid amplification tests.Analytical and bioanalytical chemistry · 2026Review
- Ultra-Rapid DNA Extraction Coupled with Loop-Mediated Isothermal Amplification for Point-of-Care Detection of Trypanosoma cruzi Infection in Heparinized Blood and Dried Blood Spots.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Extraction-free LAMP-LFD assay for early diagnosis of congenital Chagas disease directly from neonatal dried blood spots.Frontiers in cellular and infection microbiology · 2026Article
- A specific, stable, and accessible LAMP assay targeting the HSP70 gene ofMicrobiology spectrum · 2025Article
- Molecular detection and identification of Trichobilharzia: development of a LAMP, qPCR, and multiplex PCR toolkit.Parasites & vectors · 2025Article
- Diagnosing Chagas in Pregnancy and Childhood: What's Old and New.Clinics in laboratory medicine · 2025Review
- Genomics in parasitology: Accomplishments and challengesBiomedica : revista del Instituto Nacional de Salud · 2024Article
- A guide for the generation of repositories of clinical samples for research on Chagas disease.PLoS neglected tropical diseases · 2024Article
- Development of a Colorimetric Loop-Mediated Isothermal Amplification Assay for the Detection ofDiagnostics (Basel, Switzerland) · 2024Article
- Validation of a Loop-Mediated Isothermal Amplification-Based Kit for the Detection ofMicroorganisms · 2024Article
- Article
- Single nucleotide polymorphism-based visual identification ofFrontiers in plant science · 2024Article
- Combination of ultra-rapid DNA purification (PURE) and loop-mediated isothermal amplification (LAMP) for rapid detection of Trypanosoma cruzi DNA in dried blood spots.PLoS neglected tropical diseases · 2023Article
- Clinical use of molecular methods forFrontiers in parasitology · 2023Article
- Laboratory diagnosis ofFrontiers in parasitology · 2023Review
- Unveiling challenges in real-time PCR strategies for detecting treatment failure: observations from clinical trials on chronic Chagas disease.Frontiers in parasitology · 2023Review
Corrections and comments
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Authors and funding
14 authors at 4 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This study aimed to assess analytical parameters of a prototype LAMP kit that was designed for detection of Trypanosoma cruzi DNA in human blood. The prototype is based on the amplification of the highly repetitive satellite sequence of T.cruzi in microtubes containing dried reagents on the inside of the caps. The reaction is carried out at 65°C during 40 minutes. Calcein allows direct detection of amplified products with the naked eye. Inclusivity and selectivity were tested in purified DNA from Trypanosoma cruzi stocks belonging to the six discrete typing units (DTUs), in DNA from other protozoan parasites and in human DNA. Analytical sensitivity was estimated in serial dilutions of DNA samples from Sylvio X10 (Tc I) and CL Brener (Tc VI) stocks, as well as from EDTA-treated or heparinized blood samples spiked with known amounts of cultured epimastigotes (CL Brener). LAMP sensitivity was compared after DNA extraction using commercial fiberglass columns or after "Boil & Spin" rapid preparation. Moreover, the same DNA and EDTA-blood spiked samples were subjected to standardized qPCR based on the satellite DNA sequence for comparative purposes. A panel of peripheral blood specimens belonging to Chagas disease patients, including acute, congenital, chronic and reactivated cases (N = 23), as well as seronegative controls (N = 10) were evaluated by LAMP in comparison to qPCR. LAMP was able to amplify DNAs from T. cruzi stocks representative of the six DTUs, whereas it did not amplify DNAs from Leishmania sp, T. brucei sp, T. rangeli KPN+ and KPN-, P. falciparum and non-infected human DNA. Analytical sensitivity was 1x10-2 fg/μL of both CL Brener and Sylvio X10 DNAs, whereas qPCR detected up to 1x 10-1 fg/μL of CL Brener DNA and 1 fg/μl of Sylvio X10 DNA. LAMP detected 1x10-2 parasite equivalents/mL in spiked EDTA blood and 1x10-1 par.eq/mL in spiked heparinized blood using fiberglass columns for DNA extraction, whereas qPCR detected 1x10-2 par.eq./mL in EDTA blood. Boil & Spin extraction allowed detection of 1x10-2 par.eq /mL in spiked EDTA blood and 1 par.eq/ml in heparinized blood. LAMP was able to detect T.cruzi infection in peripheral blood samples collected from well-characterised seropositive patients, including acute, congenital, chronic and reactivated Chagas disease. To our knowledge, this is the first report of a prototype LAMP kit with appropriate analytical sensitivity for diagnosis of Chagas disease patients, and potentially useful for monitoring treatment response.
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