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Development and Evaluation of a Three-Dimensional Printer-Based DNA Extraction Method Coupled to Loop Mediated Isothermal Amplification for Point-of-Care Diagnosis of Congenital Chagas Disease in Endemic Regions.
- Source :
-
The Journal of molecular diagnostics : JMD [J Mol Diagn] 2021 Apr; Vol. 23 (4), pp. 389-398. Date of Electronic Publication: 2020 Dec 30. - Publication Year :
- 2021
-
Abstract
- Vertical transmission of Trypanosomacruzi is the cause of congenital Chagas disease, a re-emerging infectious disease that affects endemic and nonendemic regions alike. An early diagnosis is crucial because prompt treatment achieves a high cure rate, precluding evolution to symptomatic chronic Chagas disease. However, early diagnosis involves low-sensitive parasitologic assays, making necessary serologic confirmation after 9 months of life. With the aim of implementing early diagnostic strategies suitable for minimally equipped laboratories, a T. cruzi-loop-mediated isothermal amplification (LAMP) prototype was coupled with an automated DNA-extraction device repurposed from a three-dimensional printer (PrintrLab). The whole process takes <3 hours to yield a result, with an analytical sensitivity of 0.1 to 2 parasite equivalents per milliliter, depending on the T. cruzi strain. Twenty-five blood samples from neonates born to seropositive mothers were tested blindly. In comparison to quantitative real-time PCR, the PrintrLab-LAMP dual strategy showed high agreement, while both molecular-based methodologies yielded optimal sensitivity and specificity with respect to microscopy-based diagnosis of congenital Chagas disease. PrintrLab-LAMP detected all 10 congenitally transmitted T. cruzi infections, showing promise for point-of-care early diagnosis of congenital Chagas disease.<br /> (Copyright © 2021 Association for Molecular Pathology and American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Bolivia epidemiology
Chagas Disease epidemiology
Chagas Disease parasitology
DNA, Protozoan blood
Diagnostic Tests, Routine methods
Early Diagnosis
Female
Humans
Infant
Infant, Newborn
Infant, Newborn, Diseases blood
Infant, Newborn, Diseases epidemiology
Infant, Newborn, Diseases parasitology
Real-Time Polymerase Chain Reaction methods
Sensitivity and Specificity
Chagas Disease diagnosis
Chagas Disease transmission
DNA, Protozoan genetics
DNA, Protozoan isolation & purification
Endemic Diseases
Infant, Newborn, Diseases diagnosis
Infectious Disease Transmission, Vertical
Molecular Diagnostic Techniques methods
Nucleic Acid Amplification Techniques methods
Point-of-Care Systems
Trypanosoma cruzi genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1943-7811
- Volume :
- 23
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- The Journal of molecular diagnostics : JMD
- Publication Type :
- Academic Journal
- Accession number :
- 33387697
- Full Text :
- https://doi.org/10.1016/j.jmoldx.2020.12.007