Feriel Bouziane1, Rachida Allem2, Mohammed Sebaihia1, Sylvain Kumanski3, Faiza Mougari4, Wladimir Sougakoff5, Laurent Raskine3, Djamel Yala6, Emmanuelle Cambau7. 1. Laboratoire de Biologie Moléculaire, Génomique et Bio-Informatique-Département de Biologie, Faculté des Sciences, Université Hassiba Ben Bouali, Chlef, Algeria. 2. Laboratoire de Bio Ressources Naturelles, Département de Biologie, Faculté des Sciences, Université Hassiba Ben Bouali, Chlef, Algeria. 3. AP-HP, Laboratoire de Bactériologie, Centre National de Référence Des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, GH Lariboisière-Fernand Widal, Paris, France. 4. AP-HP, Laboratoire de Bactériologie, Centre National de Référence Des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, GH Lariboisière-Fernand Widal, Paris, France; Iame, UMR 1137, INSERM, Université Paris Diderot, Sorbonne Paris Cité, Paris, France. 5. AP-HP, Laboratoire de Bactériologie-Hygiène, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, GH Pitié-Salpêtrière, Paris, France. 6. Laboratoire National de Référence pour la Tuberculose et Mycobactéries, Institut Pasteur, Alger, Algeria. 7. AP-HP, Laboratoire de Bactériologie, Centre National de Référence Des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, GH Lariboisière-Fernand Widal, Paris, France; Iame, UMR 1137, INSERM, Université Paris Diderot, Sorbonne Paris Cité, Paris, France. Electronic address: Emmanuelle.cambau@aphp.fr.
Abstract
OBJECTIVES: To characterise the genotypes of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) isolated in Algeria, where there is a low MDR-MTB incidence rate. METHODS: Ten MDR isolates and one resistant to isoniazid were investigated by PCR-Sanger sequencing for 10 loci involved in resistance. Amplicon-based next generation sequencing (NGS) of 15 loci was additionally performed on isolates harbouring novel mutations. RESULTS: Sanger and amplicon-NGS provided the same results as with GenoType kits. Mutations known to be associated with resistance were described for most isolates: rpoB S531L in seven of 10 rifampicin-R MTB isolates, katG S315T in nine of 11 isoniazid-R, and promoter inhA c-15t in three of 11, embB M306V or M306I in two of two ethambutol-R, rpsL K43R in four of eight or rrs a514c associated with gidB L16R in streptomycin-R, gyrA A90V in the ofloxacin-R pre-XDR isolate. New and rare mutations were also described in rpoB (deletion 512-513-514), katG (S315R, M126I/ R496L), gidB (V124G, E92A, V139A, G37V), and gyrA (P8A). Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) profiles were similar for three isolates (lineage Cameroon), indicating a possible clonal diffusion in epidemiologically unrelated patients. CONCLUSIONS: Resistant MTB isolates in Algeria harbour resistance genotypes similar to other countries, but some rare patterns may result from selection and transmission processes inherent to the country.
OBJECTIVES: To characterise the genotypes of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) isolated in Algeria, where there is a low MDR-MTB incidence rate. METHODS: Ten MDR isolates and one resistant to isoniazid were investigated by PCR-Sanger sequencing for 10 loci involved in resistance. Amplicon-based next generation sequencing (NGS) of 15 loci was additionally performed on isolates harbouring novel mutations. RESULTS: Sanger and amplicon-NGS provided the same results as with GenoType kits. Mutations known to be associated with resistance were described for most isolates: rpoB S531L in seven of 10 rifampicin-R MTB isolates, katG S315T in nine of 11 isoniazid-R, and promoter inhA c-15t in three of 11, embB M306V or M306I in two of two ethambutol-R, rpsL K43R in four of eight or rrs a514c associated with gidB L16R in streptomycin-R, gyrA A90V in the ofloxacin-R pre-XDR isolate. New and rare mutations were also described in rpoB (deletion 512-513-514), katG (S315R, M126I/ R496L), gidB (V124G, E92A, V139A, G37V), and gyrA (P8A). Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) profiles were similar for three isolates (lineage Cameroon), indicating a possible clonal diffusion in epidemiologically unrelated patients. CONCLUSIONS: Resistant MTB isolates in Algeria harbour resistance genotypes similar to other countries, but some rare patterns may result from selection and transmission processes inherent to the country.
Authors: Deisy M G C Rocha; Carlos Magalhães; Baltazar Cá; Angelica Ramos; Teresa Carvalho; Iñaki Comas; João Tiago Guimarães; Helder Novais Bastos; Margarida Saraiva; Nuno S Osório Journal: Front Microbiol Date: 2021-06-11 Impact factor: 5.640