Abstract
BackgroundMycobacterial interspersed repetitive units - variable number of tandem repeats (MIRU-VNTR) genotyping is a powerful tool for unraveling clonally complex Mycobacterium tuberculosis (MTB) strains and detection of transmission patterns. Using MIRU-VNTR, MTB genotypes and their transmission patterns among patients with new and active pulmonary tuberculosis (PTB) in Kawempe municipality in Kampala, Uganda was determined.ResultsMIRU-VNTR genotyping was performed by PCR-amplification of 15 MTB-MIRU loci from 113 cultured specimens from 113 PTB patients (one culture sample per patient). To determine lineages, the genotypes were entered into the MIRU-VNTRplus database [http://www.miru-vntrplus.org/] as numerical codes corresponding to the number of alleles at each locus. Ten different lineages were obtained: Uganda II (40% of specimens), Uganda I (14%), LAM (6%), Delhi/CAS (3%), Haarlem (3%), Beijing (3%), Cameroon (3%), EAI (2%), TUR (2%) and S (1%). Uganda I and Uganda II were the most predominant genotypes. Genotypes for 29 isolates (26%) did not match any strain in the database and were considered unique. There was high diversity of MIRU-VNTR genotypes, with a total of 94 distinct patterns. Thirty four isolates grouped into 15 distinct clusters each with two to four isolates. Eight households had similar MTB strains for both index and contact cases, indicating possible transmission.ConclusionMIRU-VNTR genotyping revealed high MTB strain diversity with low clustering in Kawempe municipality. The technique has a high discriminatory power for genotyping MTB strains in Uganda.
Highlights
Mycobacterial interspersed repetitive units - variable number of tandem repeats (MIRU-VNTR)genotyping is a powerful tool for unraveling clonally complex Mycobacterium tuberculosis (MTB) strains and detection of transmission patterns
The technique has a high discriminatory power for genotyping MTB strains in Uganda
Sputum samples were collected from patients with at least one positive culture for MTB, who were previously enrolled in the Kawempe Community Household Contact Study, an ongoing epidemiological study in Kampala, Uganda, from which several papers have been published [12,13,14,15]
Summary
Genotyping is a powerful tool for unraveling clonally complex Mycobacterium tuberculosis (MTB) strains and detection of transmission patterns. Using MIRU-VNTR, MTB genotypes and their transmission patterns among patients with new and active pulmonary tuberculosis (PTB) in Kawempe municipality in Kampala, Uganda was determined. Tuberculosis (TB) is a leading cause of morbidity and mortality throughout sub-Saharan Africa, and Uganda ranks sixteenth among countries with the highest burden of disease [1]. Co-infection with HIV/AIDS and the emergence of multi-drug resistant (MDR) Mycobacterium tuberculosis (MTB) strains have made TB a major public health problem [2]. HIV infected patients [3]. TB prevalence in Uganda is believed to be higher than reported due to lack of sufficient healthcare; many people are not aware that they are infected with MTB and this has led to low levels of diagnosis and treatment [4]. Uganda has one of the lowest TB cure rates (32%) and high drug default rate [1], which may lead to an increase in drug resistance mutations.
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