학술논문

Evaluation of 16S rRNA qPCR for detection of Mycobacterium lepraeDNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases
Document Type
Article
Source
Pathogens and Global Health; February 2018, Vol. 112 Issue: 2 p72-78, 7p
Subject
Language
ISSN
20477724; 20477732
Abstract
AbstractMycobacterium lepraebacilli are mainly transmitted by the dissemination of nasal aerosols from multibacillary (MB) patients to susceptible individuals through inhalation. The upper respiratory tract represents the main entry and exit routes of M. leprae. Therefore, this study aimed to evaluate the sensitivity and specificity of real-time quantitative polymerase chain reaction (qPCR) in detecting M. lepraein nasal secretion (NS) and skin biopsy (SB) samples from MB and paucibacillary (PB) cases. Fifty-four NS samples were obtained from leprosy patients at the Dona Libânia National Reference Centre for Sanitary Dermatology in Ceará, Brazil. Among them, 19 MB cases provided both NS and SB samples. Bacilloscopy index assays were conducted and qPCR amplification was performed using specific primers for M. leprae16S rRNA gene, generating a 124-bp fragment. Primer specificity was verified by determining the amplicon melting temperature (Tm = 79.5 °C) and detection limit of qPCR was 20 fg of M. lepraeDNA. Results were positive for 89.7 and 73.3% of NS samples from MB and PB cases, respectively. SB samples from MB patients were 100% positive. The number of bacilli detected in NS samples were 1.39 × 103–8.02 × 105, and in SB samples from MB patients were 1.87 × 103–1.50 × 106. Therefore, qPCR assays using SYBR Green targeting M. leprae16S rRNA region can be employed in detecting M. lepraein nasal swabs from leprosy patients, validating this method for epidemiological studies aiming to identify healthy carriers among household contacts or within populations of an endemic area.