Benson R Kidenya1, Stephen E Mshana2, Daniel W Fitzgerald3, Oksana Ocheretina3. 1. Department of Biochemistry and Molecular Biology, Catholic University of Health and Allied Sciences, P.O. Box 1464, Mwanza, Tanzania. Electronic address: benkidenya@bugando.ac.tz. 2. Department of Medical Microbiology/Immunology, Catholic University of Health and Allied Sciences, Mwanza, Tanzania. 3. Center for Global Health, Division of Infectious Diseases, Weill Cornell Medical College, New York, NY, USA.
Abstract
BACKGROUND: Drug resistant Tuberculosis (TB) is considered a global public health threat. Whole-genome sequencing (WGS) is a new technology for tuberculosis (TB) diagnostics and is capable of providing rapid drug resistance profiles and genotypes for epidemiologic surveillance. Therefore, we used WGS to determine genotypic drug resistance profiles and genetic diversity of drug resistant Mycobacterium tuberculosis isolates from Mwanza, North-western Tanzania. METHODS: A cross-sectional study was conducted at the Bugando Medical Center (BMC) from September 2014 to June 2015. Consecutively, smear-positive newly diagnosed TB patients aged ≥18 years were enrolled. Sputum samples were cultured on Löwenstein-Jensen (LJ) slants. Mycobacterial genomic DNA was extracted for WGS to determine drug resistant mutations for first and second line drugs as well as the spoligotypes. RESULTS: A total of 78 newly diagnosed patients with pulmonary TB with a median age of 37 [IQR: 30-46] years were enrolled. Of these, 57.8% (45/74) were males and 34.6% (27/78) were HIV positive. Mycobacterium tuberculosis genomic DNA for WGS was obtained from isolates in 74 (94.9%) patients. Of the 74 isolates, six (8.1%) isolates harbored mutations for resistance to at least one drug. The resistance to the drugs was isoniazid 3/74 (4.1%), rifampicin mono-resistant 2/74 (2.7%), ethambutol 2/74 (2.7%) and streptomycin 1/74 (1.4%). None was isoniazid mono-resistant. Of the 74 only one (1.4%) patient had MDR-TB. The resistance to ethionamide, the second line drug, was detected in one patient (1.4%). None was resistant to pyrazinamide, fluoroquinolones, kanamycin, amikacin, or capreomycin. The mutations detected were mabA-inhA promoter region C(-15)T and katG Ser513Thr for isoniazid; rpoB His526Leu and rpoB Ser531Leu for rifampicin; embB Met306Val and embB Met306Ile for ethambutol; rpsL Lys43Arg for streptomycin; and mabA-inhA promoter region C(-15)T for ethionamide. The spoligotypes of the drug resistant Mycobacterium tuberculosis were distinct to all six isolates and belonged to T1, T2, T3-ETH, CAS1-DELHI, EAI5 and LAM11-ZWE lineages. CONCLUSION: The genetic drug resistance profile of Mycobacterium tuberculosis isolates from North-western Tanzania comprises of the common previously reported mutations. The prevalence of resistance to first and second line drugs including MDR-TB is low. Six drug resistant strains exhibited different spoligotypes, suggesting limited transmission of drug resistant strains in the region.
BACKGROUND:Drug resistant Tuberculosis (TB) is considered a global public health threat. Whole-genome sequencing (WGS) is a new technology for tuberculosis (TB) diagnostics and is capable of providing rapid drug resistance profiles and genotypes for epidemiologic surveillance. Therefore, we used WGS to determine genotypic drug resistance profiles and genetic diversity of drug resistant Mycobacterium tuberculosis isolates from Mwanza, North-western Tanzania. METHODS: A cross-sectional study was conducted at the Bugando Medical Center (BMC) from September 2014 to June 2015. Consecutively, smear-positive newly diagnosed TB patients aged ≥18 years were enrolled. Sputum samples were cultured on Löwenstein-Jensen (LJ) slants. Mycobacterial genomic DNA was extracted for WGS to determine drug resistant mutations for first and second line drugs as well as the spoligotypes. RESULTS: A total of 78 newly diagnosed patients with pulmonary TB with a median age of 37 [IQR: 30-46] years were enrolled. Of these, 57.8% (45/74) were males and 34.6% (27/78) were HIV positive. Mycobacterium tuberculosis genomic DNA for WGS was obtained from isolates in 74 (94.9%) patients. Of the 74 isolates, six (8.1%) isolates harbored mutations for resistance to at least one drug. The resistance to the drugs was isoniazid 3/74 (4.1%), rifampicin mono-resistant 2/74 (2.7%), ethambutol 2/74 (2.7%) and streptomycin 1/74 (1.4%). None was isoniazid mono-resistant. Of the 74 only one (1.4%) patient had MDR-TB. The resistance to ethionamide, the second line drug, was detected in one patient (1.4%). None was resistant to pyrazinamide, fluoroquinolones, kanamycin, amikacin, or capreomycin. The mutations detected were mabA-inhA promoter region C(-15)T and katG Ser513Thr for isoniazid; rpoB His526Leu and rpoB Ser531Leu for rifampicin; embB Met306Val and embB Met306Ile for ethambutol; rpsL Lys43Arg for streptomycin; and mabA-inhA promoter region C(-15)T for ethionamide. The spoligotypes of the drug resistant Mycobacterium tuberculosis were distinct to all six isolates and belonged to T1, T2, T3-ETH, CAS1-DELHI, EAI5 and LAM11-ZWE lineages. CONCLUSION: The genetic drug resistance profile of Mycobacterium tuberculosis isolates from North-western Tanzania comprises of the common previously reported mutations. The prevalence of resistance to first and second line drugs including MDR-TB is low. Six drug resistant strains exhibited different spoligotypes, suggesting limited transmission of drug resistant strains in the region.
Authors: Aaron McKenna; Matthew Hanna; Eric Banks; Andrey Sivachenko; Kristian Cibulskis; Andrew Kernytsky; Kiran Garimella; David Altshuler; Stacey Gabriel; Mark Daly; Mark A DePristo Journal: Genome Res Date: 2010-07-19 Impact factor: 9.043
Authors: T M Chonde; D Basra; S G M Mfinanga; N Range; F Lwilla; R P Shirima; A van Deun; M Zignol; F G Cobelens; S M Egwaga; F van Leth Journal: Int J Tuberc Lung Dis Date: 2010-08 Impact factor: 2.373
Authors: J D van Embden; M D Cave; J T Crawford; J W Dale; K D Eisenach; B Gicquel; P Hermans; C Martin; R McAdam; T M Shinnick Journal: J Clin Microbiol Date: 1993-02 Impact factor: 5.948
Authors: Catharina C Boehme; Pamela Nabeta; Doris Hillemann; Mark P Nicol; Shubhada Shenai; Fiorella Krapp; Jenny Allen; Rasim Tahirli; Robert Blakemore; Roxana Rustomjee; Ana Milovic; Martin Jones; Sean M O'Brien; David H Persing; Sabine Ruesch-Gerdes; Eduardo Gotuzzo; Camilla Rodrigues; David Alland; Mark D Perkins Journal: N Engl J Med Date: 2010-09-01 Impact factor: 91.245
Authors: Deus Lukoye; Fred A Katabazi; Kenneth Musisi; David P Kateete; Benon B Asiimwe; Moses Okee; Moses L Joloba; Frank G J Cobelens Journal: Antimicrob Agents Chemother Date: 2014-04-28 Impact factor: 5.191
Authors: M de Vos; B Müller; S Borrell; P A Black; P D van Helden; R M Warren; S Gagneux; T C Victor Journal: Antimicrob Agents Chemother Date: 2012-12-03 Impact factor: 5.191
Authors: Willy Ssengooba; Conor J Meehan; Deus Lukoye; George William Kasule; Kenneth Musisi; Moses L Joloba; Frank G Cobelens; Bouke C de Jong Journal: Infect Genet Evol Date: 2016-02-23 Impact factor: 3.342
Authors: Timothy M Walker; Camilla L C Ip; Ruth H Harrell; Jason T Evans; Georgia Kapatai; Martin J Dedicoat; David W Eyre; Daniel J Wilson; Peter M Hawkey; Derrick W Crook; Julian Parkhill; David Harris; A Sarah Walker; Rory Bowden; Philip Monk; E Grace Smith; Tim E A Peto Journal: Lancet Infect Dis Date: 2012-11-15 Impact factor: 25.071
Authors: Peter M Mbelele; Christian Utpatel; Elingarami Sauli; Emmanuel A Mpolya; Beatrice K Mutayoba; Ivan Barilar; Viola Dreyer; Matthias Merker; Margaretha L Sariko; Buliga M Swema; Blandina T Mmbaga; Jean Gratz; Kennedy K Addo; Michel Pletschette; Stefan Niemann; Eric R Houpt; Stellah G Mpagama; Scott K Heysell Journal: JAC Antimicrob Resist Date: 2022-04-21
Authors: Bugwesa Z Katale; Peter M Mbelele; Nsiande A Lema; Susana Campino; Stephen E Mshana; Mark M Rweyemamu; Jody E Phelan; Julius D Keyyu; Mtebe Majigo; Erasto V Mbugi; Hazel M Dockrell; Taane G Clark; Mecky I Matee; Stellah Mpagama Journal: BMC Genomics Date: 2020-02-21 Impact factor: 3.969
Authors: Asma Munir; Michael T Wilson; Steven W Hardwick; Dimitri Y Chirgadze; Jonathan A R Worrall; Tom L Blundell; Amanda K Chaplin Journal: Structure Date: 2021-01-13 Impact factor: 5.006