Title :

Validation of Swab Sampling and SYBR Green-Based Real-Time PCR for the Diagnosis of Cutaneous Leishmaniasis in French Guiana

Author(s) :

Blaizot, Romain [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Simon, Stéphane [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Ginouvès, Marine [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Prévôt, Ghislaine [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Blanchet, Denis [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Ravel, Christophe [Auteur]
Biologie, Génétique et Pathologie des Pathogènes Eucaryotes [MIVEGEC-BioGEPPE]
Centre National de Référence des Leishmanioses [CHRU Montpellier] [CNR-L]
Couppié, Pierre [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Demar, Magalie Pierre [Auteur]
Biomes Tropicaux et Immuno-Pathophysiologie = Tropical Biome and ImmunoPhysiopathology [Lille] [TBIP]
Nabet, Cécile [Auteur]
Institut Pierre Louis d'Epidémiologie et de Santé Publique [iPLESP]
CHU Pitié-Salpêtrière [AP-HP]

Journal title :

Journal of Clinical Microbiology

Publisher :

American Society for Microbiology

Publication date :

2021-01-21

ISSN :

0095-1137

English keyword(s) :

cutaneous leishmaniasis
diagnostic test
PCR
neglected tropical disease

HAL domain(s) :

Sciences du Vivant [q-bio]/Immunologie

English abstract : [en]

Recent studies have highlighted the interest in noninvasive sampling procedures coupled with real-time PCR methods for the detection of Leishmania species in South America. In French Guiana, the sampling method still relied ...
Show more >Recent studies have highlighted the interest in noninvasive sampling procedures coupled with real-time PCR methods for the detection of Leishmania species in South America. In French Guiana, the sampling method still relied on skin biopsies. Noninvasive protocols should be tested on a large annual cohort to improve routine laboratory diagnosis of cutaneous leishmaniasis. Therefore, we evaluated the performance of a new Leishmania detection and species identification protocol involving cotton swabs and SYBR green-based real-time PCR of the Hsp70 gene, coupled with Sanger sequencing. Between May 2017 and May 2018, 145 patients with ulcerated lesions compatible with cutaneous leishmaniasis were included in the study at the Cayenne Hospital and its remote health centers. Each patient underwent scrapings for a smear, skin biopsies for parasite culture and PCR-restriction fragment length polymorphism (RFLP) (RNA polymerase II), and sampling with a cotton swab for SYBR green-based PCR. The most accurate diagnostic test was the SYBR green-based PCR on swab samples, showing 98% sensitivity. The mean PCR cycle threshold (CT) was 24.4 (minimum CT, 17; maximum CT, 36) and was <35 in 97.6% of samples. All samples positive by SYBR green-based real-time PCR were successfully identified at the species level by DNA sequencing. This new method should be considered for routine diagnosis of cutaneous leishmaniasis in South America and especially for remote areas, since noninvasive collection tools are easier to use and require fewer precautions for transportation.Show less >

Language :

Anglais

Peer reviewed article :

Oui

Audience :

Internationale

Popular science :

Non

Collections :

• Centre d'Infection et d'Immunité de Lille (CIIL) - U1019 - UMR 9017

Source :

Harvested from HAL