Fluorescent Hybridization of Mycobacterium leprae in Skin Samples Collected in Burkina Faso

Abstract

Leprosy is caused by Mycobacterium leprae, and it remains underdiagnosed in Burkina Faso. We investigated the use of fluorescent in situ hybridization (FISH) for detecting M. leprae in 27 skin samples (skin biopsy samples, slit skin samples, and skin lesion swabs) collected from 21 patients from Burkina Faso and three from Côte d’Ivoire who were suspected of having cutaneous leprosy. In all seven Ziehl-Neelsen-positive skin samples (four skin biopsy samples and three skin swabs collected from the same patient), FISH specifically identified M. leprae, including one FISH-positive skin biopsy sample that remained negative after testing with PCR targeting the rpoB gene and with the GenoType LepraeDR assay. Twenty other skin samples and three negative controls all remained negative for Ziehl-Neelsen staining, FISH, and rpoB PCR. These data indicate the usefulness of a microscopic examination of skin samples after FISH for first-line diagnosis of cutaneous leprosy. Accordingly, FISH represents a potentially useful point-of-care test for the diagnosis of cutaneous leprosy.

Whole genome sequence of bacteremic Clostridium tertium in a World War I soldier, 1914

“…Briefly, the dental pulp of individual SQ517 was fixed for three hours in Sandison's rehydration solution (aqueous formaldehyde 1%, 96% ethanol and 5% aqueous sodium carbonate) ( Collini et al., 2014 ) and smeared on microscopic slade using Cytospin (ThermoFisher, Illkirch, France). In situ hybridization was performed with a hybridiser (Dako, Les Ulis, France) at 65 °C for 10 min and then at 37° overnight, following a previously reported protocol ( Millogo et al., 2020 ). Dental pulp of the individual SQ536, which remained negative in culture for C. tertium was used as a negative control to assess the probe specificity.…”

Fecal Excretion of Mycobacterium leprae, Burkina Faso

“…Leprosy caused by Mycobacterium leprae remains endemic in Burkina Faso, a West Africa country with a level of disability 2 of 31.2% among new patient cases ( 1 ). Laboratory diagnosis of leprosy is determined by observation of acid-fast bacilli after microscopic examination of a Ziehl-Neelsen–stained nasal smears and cutaneous lesions ( 1 ). Recently, fluorescence in situ hybridization (FISH) was introduced as a complementary approach to increase the specificity of microscopic observations ( 1 , 2 ).…”

“…Laboratory diagnosis of leprosy is determined by observation of acid-fast bacilli after microscopic examination of a Ziehl-Neelsen–stained nasal smears and cutaneous lesions ( 1 ). Recently, fluorescence in situ hybridization (FISH) was introduced as a complementary approach to increase the specificity of microscopic observations ( 1 , 2 ). We report on the specific microscopic detection of M. leprae in the stool specimen of a patient in Burkina Faso.…”

“…All samples were microscopically examined after Ziehl-Neelsen staining and revealed acid-fast bacilli in all 5 samples. Acid-fast bacilli were further identified as M. leprae by partial PCR amplification sequencing of the rpoB gene using a validated protocol ( 1 ).…”

Confirming Autochthonous Buruli Ulcer Cases in Burkina Faso, West Africa

“…The ulcers were classified as WHO category II and WHO category III (Figure 2). DNA extracted using the QIAGEN Kit (Qiagen, Hilden, Germany) combined with the glass powder treatment, heating at 56 C for 2 h, sonication for 30 min, and automatic elution with an EZ1 DNA tissue kit (Qiagen) as reported previously 5 was incorporated into three real-time quantitative polymerase chain reactions targeting Mycobacterium ulcerans insertion sequences IS2404, IS2606 and the KR-B gene as described by Fyfe. 6 These manipulations were carried out at the Marseille Institut Hospitalier Universitaire (Marseille, France) in the presence of negative controls (Supplemental Table 1).…”