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Título
Detection of Schistosoma mansoni-derived
DNA in human urine samples by loopmediated
isothermal amplification (LAMP)
Autor(es)
Palabras clave
Parasitic diseases
Urine
Schistosomiasis
Diagnostic medicine
DNA
Clasificación UNESCO
3207.12 Parasitología
Fecha de publicación
2019-03-26
Editor
Public Library of Science (New York)
Citación
Fernández-Soto P, Gandasegui J,
Carranza Rodríguez C, Pérez-Arellano JL, Crego-
Vicente B, García-Bernalt Diego J, et al. (2019)
Detection of Schistosoma mansoni-derived DNA in
human urine samples by loop-mediated isothermal
amplification (LAMP). PLoS ONE 14(3): e0214125.
https://doi.org/10.1371/journal.pone.0214125
Resumen
[EN]Background
Schistosoma mansoni is the main species causing hepatic and intestinal schistosomiasis in
Sub-Saharan Africa, and it is the only species in South America. Adult stages of the parasite
reside in the mesenteric venous plexus of infected hosts, and eggs are shed in feces. Collecting
patient stool samples for S. mansoni diagnostic purposes is difficult in large-scale
field trials. Urine samples would be an alternative approach for molecular S. mansoni detection
since they have several advantages over stool samples, including better handling, management
and storage. Additionally, loop-mediated isothermal amplification (LAMP)
technology is a powerful molecular diagnostic tool for infectious diseases, particularly under
field conditions in developing countries. The present study aimed to assess the effectiveness
of our previously developed LAMP assay (SmMIT-LAMP) for S. mansoni-specific
detection in clinical urine samples.
Methodology/Principal findings
The sensitivity of SmMIT-LAMP in urine was established in simulated fresh human urine
samples artificially spiked with genomic DNA from S. mansoni. LAMP for 120 min instead of
60 min improved the sensitivity, reaching values of 0.01 fg/μL. A set of well-defined frozen
stored human urine samples collected from Sub-Saharan immigrant patients was selected
from a biobank to evaluate the diagnostic validity of SmMIT-LAMP. The set included urine
samples from patients with microscopy-confirmed infections with S. mansoni, S. haematobium
and other nonschistosome parasites, as well as urine samples from patients with
microscopy-negative eosinophilia without a confirmed diagnosis. The SmMIT-LAMP was
incubated for 60 and 120 min. A longer incubation time was shown to increase the LAMPpositive
results in patient urine samples. We also tested urine samples from mice experimentally
infected with S. mansoni, and LAMP-positive results were obtained from the third week after infection. A real-time LAMP assay was also performed with three individual urine
samples.
Conclusions/Significance
The SmMIT-LAMP could effectively detect S. mansoni DNA in mouse urine samples and
produced promising results for human clinical samples. The detection of S. mansoni DNA in
mouse urine samples from the third week after infection indicates that early diagnosis of
active S. mansoni infection is possible using urine as a source of DNA. Further studies are
still needed, but our method could be used as a promising molecular tool applicable to urine
samples to diagnose human intestinal schistosomiasis caused by S. mansoni.
URI
ISSN
1932-6203
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