Abstract
BackgroundTuberculosis (TB) in both animals and humans is caused by Mycobacterium tuberculosis complex (MTBC) primarily transmitted by inhalation of aerosolized droplets containing the organism. Multi-drug resistance (MDR) and extensive drug resistance (XDR) are evolutionary features of Mycobacterium tuberculosis to subvert the antibiotic regimes in place. The heavy burden of TB worsened by HIV endemic in South Africa motivated for the investigation of MTBC prevalence among TB patients in Port Elizabeth and the amplification and sequencing of the DNA amplicons known to confer resistance to TB drugs.MethodsThree thousand eight hundred and ten (3810) sputum specimens were processed and DNA was isolated from sputum specimens collected from different hospitals and health care places in the Eastern Cape Province, South Africa. DNA was amplified using the Seeplex® MTB Nested ACE detection assay. The agar-dilution proportion method was used to perform drug-sensitivity testing using 7H10 Middlebrook medium. Target genes known to confer resistance to first and second-line drugs were amplified and the amplicons sequenced.ResultsOne hundred and ninety (5%) DNA samples tested positive for MTBC and from the resistant profiles of the 190 positive samples, we noted that multidrug-resistant TB was identified in 189 (99.5%) with 190 (100%) patients infected with MTB resistant to isoniazid and 189 (99.5%) having MTB resistant to rifampicin. Other percentages of drug resistance observed including 40% pre-XDR and 60% of XDR.ConclusionThis study provides valuable data on the different kinds of mutations occurring at various target loci in resistant MTBC strains isolated from samples obtained from the Eastern Cape Province. The results obtained reveal a high incidence of MDR amongst the positive samples from Eastern Cape Province, South Africa.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2334-14-479) contains supplementary material, which is available to authorized users.
Highlights
Tuberculosis (TB) in both animals and humans is caused by Mycobacterium tuberculosis complex (MTBC) primarily transmitted by inhalation of aerosolized droplets containing the organism
Amplification of Seeplex Deoxy-Ribonucleic Acid (DNA) samples results One hundred and ninety (190) DNA samples out of the three thousand eight hundred and ten (3810) specimens were used in the study
The main strength of the Seeplex® MTB Nested ACE detection assay is that it uses multi-target Polymerase Chain Reaction (PCR) (IS6110 and mpb64) for the specific detection of MTBC only; this prevents false positive results that are caused by other mycobacteria
Summary
Tuberculosis (TB) in both animals and humans is caused by Mycobacterium tuberculosis complex (MTBC) primarily transmitted by inhalation of aerosolized droplets containing the organism. Multi-drug resistance (MDR) and extensive drug resistance (XDR) are evolutionary features of Mycobacterium tuberculosis to subvert the antibiotic regimes in place. The heavy burden of TB worsened by HIV endemic in South Africa motivated for the investigation of MTBC prevalence among TB patients in Port Elizabeth and the amplification and sequencing of the DNA amplicons known to confer resistance to TB drugs. The Mycobacterium tuberculosis Complex (MTBC) consists of nine bacterial species that cause tuberculosis (TB) in mammals, including human beings [1]. Resistance of the organisms to TB drugs is a major public health problem that threatens the progress made in TB control worldwide. There are several factors that contribute to the development of MDR-TB, such as poor adherence of patients to first line anti-TB drugs, dosage and duration of treatment, inappropriate treatment regiments and noncompliance to national guidelines and TB protocol by TB clinicians [11]
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