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Open Access 01-12-2019 | Leprosy | Technical advance

Development of a combined RLEP/16S rRNA (RT) qPCR assay for the detection of viable M. leprae from nasal swab samples

Authors: Marcus Beissner, Anna Woestemeier, Malkin Saar, Kossi Badziklou, Issaka Maman, Charlotte Amedifou, Magdalena Wagner, Franz X. Wiedemann, Komi Amekuse, Basile Kobara, Karl-Heinz Herbinger, Abiba Banla Kere, Thomas Löscher, Gisela Bretzel

Published in: BMC Infectious Diseases | Issue 1/2019

Abstract

Background

Leprosy continues to be a health problem in endemic areas. More than 200,000 new cases of leprosy per year suggest that transmission of the disease is still ongoing, presumably as airborne infection through nasal droplets. Late diagnosis supports continued transmission and increases the individual risk for functional disabilities. Laboratory tools are considered beneficial to facilitate early detection and clinical assessment of cases. The aim of this study was to validate molecular tools allowing detection, quantification and assessment of viability of M. leprae from nasal swab samples which are easy to obtain without the need of any invasive procedures.

Methods

Validation of two real-time PCRs detecting M. leprae DNA (RLEP qPCR) and RNA (16S rRNA RT qPCR) was conducted on “must not detect”/“must detect” samples and 160 pre-treatment nasal swab samples from 20 clinically diagnosed multibacillary (MB) leprosy patients from Togo.

Results

Both assays were 100% M. leprae specific and showed analytical sensitivities of three templates each. Out of 20 clinically diagnosed MB leprosy patients, 15 (75.0%) had a positive RLEP qPCR result from nasal swab samples. The 16S rRNA RT qPCR detected viable bacilli in nasal swab samples of ten out of these 15 RLEP positive patients (66.7%).

Conclusion

The combined RLEP/16S rRNA (RT) qPCR assay provides a sensitive and specific tool to determine the bacterial load and viability of M. leprae from nasal swab samples and is applicable for early diagnosis, monitoring treatment response and investigating the role of nasal carriage of M. leprae in human-to-human transmission through aerosol infection.

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Title: Development of a combined RLEP/16S rRNA (RT) qPCR assay for the detection of viable M. leprae from nasal swab samples
Authors: Marcus Beissner
Anna Woestemeier
Malkin Saar
Kossi Badziklou
Issaka Maman
Charlotte Amedifou
Magdalena Wagner
Franz X. Wiedemann
Komi Amekuse
Basile Kobara
Karl-Heinz Herbinger
Abiba Banla Kere
Thomas Löscher
Gisela Bretzel
Publication date: 01-12-2019
Publisher: BioMed Central
Keyword: Leprosy
Published in: BMC Infectious Diseases / Issue 1/2019
Electronic ISSN: 1471-2334
DOI: https://doi.org/10.1186/s12879-019-4349-9

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