Agar barrier method for removal of yeast and other fungal contamination from<i>Leishmania</i>promastigotes culture

Gupta, Anil Kumar; Narayan, Shyam; Singh, Shubhankar Kumar; Thakur, Ajit Kumar; Das, Pradeep

doi:10.1179/2047773212y.0000000034

Cited by 3 publications

(1 citation statement)

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“…However, empirical on November 25, 2020 by guest http://jcm.asm.org/ Downloaded from experience from both our laboratory and the National Reference Centre of Montpellier, indicate that antifungals might hamper Leishmania growth. These empirical findings are also supported by some publications (27) . Conditions of transport probably play an important part in preventing contaminations.…”

Section: Skin Biopsies Fromsupporting

confidence: 85%

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

et al. 2021

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Recent studies have highlighted the interest of non-invasive sampling procedures coupled to real-time PCR methods for detection of Leishmania species in South America. In French Guiana, sampling method still relied on skin biopsies. Non-invasive protocols should be tested on a large annual cohort to improve routine laboratory diagnosis of Cutaneous Leishmaniasis. Therefore, we evaluated the performances of a new Leishmania detection and species identification protocol involving cotton swabs and SYBR Green real-time PCR of 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 Cayenne Hospital and its remote health centres. Each patient underwent scrapings for smear, skin biopsies for parasite culture and PCR-RFLP (RNA pol II) and cotton swabs for SYBR Green PCR. The most accurate diagnostic test was the SYBR Green PCR on swab sampling, showing a 98% sensitivity. Mean PCR cycle threshold (Ct) was of 24.4 Ct (min=17 Ct, max=36 Ct) and was inferior to 35 Ct in 97.6% of samples. All samples positive with real-time PCR SYBR Green 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, as non-invasive collection tools are easier to use and require less precautions for transportation.

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Section: Skin Biopsies Fromsupporting

confidence: 85%

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

et al. 2021

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Isolation of naturally infecting Leishmania infantum from canine samples in Novy-MacNeal-Nicolle medium prepared with defibrinated blood from different animal species

Santos

Pinho

Passos

et al. 2018

Veterinary Parasitology

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Leishmaniasis: diagnostic issues in Europe

Torpiano¹,

Pace

2015

Expert Review of Anti-infective Therapy

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Leishmaniasis is a parasitic disease with clinical presentations that vary from asymptomatic infection to cutaneous, mucocutaneous or visceral disease. Recent epidemiological studies have shown an increased prevalence in Europe largely caused by an increase in international travel, difficulty eradicating leishmanial infection in AIDS patients, and the use of immunosuppressive medications. Clinical diagnosis may be challenging, and parasitological diagnosis entails the use of invasive procedures which may be unrevealing in the immunosuppressed. A number of less invasive tests for the detection of anti-leishmanial antibodies or leishmanial antigen are available but their sensitivity and specificity may vary with the infective species and results have to be interpreted in light of the clinical presentation. The availability of polymerase chain reaction assays amplifying leishmanial genetic material has been a major step forward in improving the diagnosis of leishmanial disease and the response to treatment.

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Agar barrier method for removal of yeast and other fungal contamination fromLeishmaniapromastigotes culture

Cited by 3 publications

References 4 publications

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

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

Isolation of naturally infecting Leishmania infantum from canine samples in Novy-MacNeal-Nicolle medium prepared with defibrinated blood from different animal species

Leishmaniasis: diagnostic issues in Europe

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