MiniPCR as a portable equipment for the molecular diagnosis of american cutaneous leishmaniasis.
Acta Trop
; 243: 106926, 2023 Jul.
Article
em En
| MEDLINE
| ID: mdl-37088354
ABSTRACT
There is an urgent need to improve the diagnostic capacity of cutaneous leishmaniasis (CL) in rural health centers to improve the management of the disease in patients from remote regions where the infection is endemic. Microscopy of Giemsa-stained lesion smears is the standard-of-care diagnostic test in virtually all health centers, but its sensitivity is suboptimal (50-70%) and prone to false negative results. We evaluated the performance of a low-cost DNA extraction buffer (LAB) using a portable miniPCR™ equipment coupled with an inexpensive fluorescence viewer to detect Leishmania DNA with the naked eye or using a commercial photo app. Using ten-fold serial dilutions of Leishmania (V.) panamensis promastigotes the miniPCR-F test detected 10 parasites per µL, which was comparable to real-time PCR. Utilization of DNA from retrospective clinical samples preserved at -80 °C from Colombia (n = 28) or lesion exudate preserved in filter papers from Peru (n = 48) showed that the miniPCR-fluorescent test had a 100% sensitivity and > 90% specificity compared to real-time PCR. This study demonstrated the utility of LAB DNA extraction method for direct amplification of Leishmania using the miniPCR and reading of P51 results with the naked eye or via digital reading with a photo app. These preliminary results indicated that the miniPCR-F test workflow could be amenable to implementation in resource-limited health centers.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Leishmaniose
/
Leishmaniose Cutânea
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Leishmania
Tipo de estudo:
Diagnostic_studies
/
Observational_studies
/
Risk_factors_studies
Limite:
Humans
Idioma:
En
Revista:
Acta Trop
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Estados Unidos