Fast diagnosis of sporotrichosis caused by Sporothrix globosa, Sporothrix schenckii, and Sporothrix brasiliensis based on multiplex real-time PCR

PLoS Neglected Tropical Diseases
Mingrui ZhangFei Zhao

Abstract

The accurate diagnosis of sporotrichosis and identification at the species level are critical for public health and appropriate patient management. Compared with morphological identification methods, molecular diagnostic tests are rapid and have high sensitivity and standardized operating processes. Therefore, we designed a novel multiplex real-time polymerase chain reaction (PCR) method based on the calmodulin (CAL) gene for the identification of clinically relevant Sporothrix species: S. globosa, S. schenckii s. str., and S. brasiliensis. We evaluated the assay with clinical and spiked samples and assessed its diagnostic performance by comparing the results to those of culture and species-specific PCR. Thirty-three DNA templates were used to detect assay specificity, and three plasmids were constructed to create a standard curve and determine the limits of detection (LODs). For nucleic acid detection, the sensitivity and specificity reached 100%. The LODs were 10 copies, 10 copies and 100 copies for S. globosa, S. schenckii s. str and S. brasiliensis, respectively. For the clinical samples, the positive detection rates by culture, species-specific PCR and the multiplex real-time PCR assay were 87.9% (29/33), 39.4% (13/33), an...Continue Reading

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Citations

Oct 24, 2019·Medical Mycology·Nurul Khalisah Kamal AzamSatinee Suetrong
Oct 31, 2020·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Conchita TorielloMaría Del Rocío Reyes-Montes
Jun 16, 2021·Veterinary Research Communications·Taiza Maschio-LimaMargarete Teresa Gottardo de Almeida
Oct 24, 2021·Transboundary and Emerging Diseases·Paula Portella Della TerraAnderson Messias Rodrigues

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Methods Mentioned

BETA
biopsies
PCR
biopsy
electrophoresis

Software Mentioned

CFX Manager
Primer Express
Statistical Analysis System

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