Spoligotyping of Mycobacterium tuberculosis isolates using Luminex®-based method in Lebanon

Khaldoun Masoud; George F Araj; Lina  Reslan; Sukayna  Fadlallah; Michel Wehbe; Lina Itani; Aline Avedissian; Ghassan  Dbaibo; Antoine Saade; Guislaine Refrégier; Christophe Sola; Ghassan M Matar

doi:10.3855/jidc.12072

Authors

Khaldoun Masoud WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
George F Araj WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Lina Reslan WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Sukayna Fadlallah WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Michel Wehbe WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Lina Itani WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Aline Avedissian WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Ghassan Dbaibo WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon
Antoine Saade Ministry of Public Health, Beirut, Lebanon
Guislaine Refrégier Institute for Integrative Biology of the Cell, University Paris‐Sud, University Paris‐Saclay, France
Christophe Sola Institute for Integrative Biology of the Cell, University Paris‐Sud, University Paris‐Saclay, France
Ghassan M Matar WHO Collaborating Center for Reference and Research on Bacterial Pathogens, Center for Infectious Diseases Research (CIDR), Faculty of Medicine, American University of Beirut, Beirut, Lebanon

DOI:

https://doi.org/10.3855/jidc.12072

Keywords:

Spoligotyping, Luminex, Mycobacterium tuberculosis, Lebanon

Abstract

Introduction: Data about the genotypes of circulating Mycobacterium tuberculosis isolates (MTB) in Lebanon are scarce. This study was undertaken to reveal the spoligotypes of MTB isolates recovered from patients in Lebanon.

Methodology: MTB isolates from 49 patients living in Lebanon were recovered and identified. The samples were heat killed and subjected to DNA extraction. Spoligotyping was performed using microbeads from TB-SPOL Kit and the fluorescence intensity was measured using Luminex 200®. Generated patterns were assigned to families using the SITVIT2 international database of the Pasteur Institute of Guadeloupe and compared.

Results: The spoligotyping of the 49 MTB isolates revealed that 31 isolates belonged to Lineage 4 (Euro-American, 63.3%), 12 to Lineage 3 (East- African Indian, 24.5%), 3 to Lineage 2 (East Asian, 6%) and 2 were unknown. Over half of the genotypes (16 of 30) harbored SIT127 supposed to belong to the L4.5 sublineage. One isolate belonging to the rare Manu-Ancestor SIT523 was recovered for the first time in Lebanon, being associated with highly virulent extensively drug-resistant (XDR) MTB phenotype.

Conclusion: The application of the Spoligotyping Multiplex Luminex® method is an efficient, discriminatory and rapid method to use for first-lane genotyping of MTB isolates. Though humble numbers were tested, this study is one of the first to describe the genomic diversity and epidemiology of MTB isolates of Lebanon, and suggests an increasing prevalence of SIT127 in the country.