Research Article ISSN 2639-9458

Microbiology & Infectious Diseases

Occurrence and Frequency of Gene Mutations Associated with

Rifampicin (RIF) and Isoniazid (INH) Resistance from Multidrug Resistant
Mycobacterium Tuberculosis (MDRTB) in Lagos, Nigeria
Raheem TY1*, Iwalokun BA1, Fowora MA1, Osuolale KA2 and Adesesan AA3

1
Molecular Biology and Biotechnology Department, Nigerian
Institute of Medical Research Yaba Lagos. *
Correspondence:
2
Grant, Monitoring and Evaluation Unit (Biostatistics), Nigerian Raheem TY, Molecular Biology and Biotechnology Department,
Nigerian Institute of Medical Research Yaba Lagos.
Institute of Medical Research.
Received: 09 Dec 2023; Accepted: 14 Jan 2024; Published: 20 Jan 2024
3
Centre for TB Research, Nigerian Institute of Medical Research
Yaba, Lagos.

Citation: Raheem TY, Iwalokun BA, Fowora MA, et al. Occurrence and Frequency of Gene Mutations Associated with Rifampicin
(RIF) and Isoniazid (INH) Resistance from Multidrug Resistant Mycobacterium Tuberculosis (MDRTB) in Lagos, Nigeria. Microbiol
Infect Dis. 2024; 8(1): 1-6.

ABSTRACT
Background: Despite the tremendous improvements in the diagnosis and treatment of tuberculosis, pulmonary tuberculosis (PTB)
remains a leading cause of morbidity and mortality in adults and children worldwide. The World Health Organization (WHO)
recommends detection of MDR TB using bacteriological confirmation of TB and testing for drug resistance using rapid molecular tests,
culture methods or sequencing technologies. M. tuberculosis strains resistant to multiple anti-TB drugs are becoming increasingly
common. Different independent mutations of rpoB, katG and InhA genes encoding either the drug target or the enzymes involved in
drug activation have been found to be one of the strategies responsible for resistance to Rifampicin and Isoniazid.
Methods: This cross-sectional, multicentre study was conducted on MDRTB isolates collected from pulmonary TB patients in
Lagos, Nigeria from May 2012 to October 2016. After informed consent, structured questionnaires were administered to obtain
sociodemographic data. Sputum samples were collected and processed for microscopy and culture using Lowenstein-Jensen medium.
Isolates were identified by biochemical and molecular methods and drug susceptibility testing was performed using MIC, proportion
and line probe Assay methods.
Objectives: The study investigated the occurrence and frequency of gene mutations associated with rifampicin (RIF) and isoniazid
(INH) resistance from the isolated MDRTB and assessed whether all the M. tuberculosis which elicited MDRTB phenotype had
mutation markers.
Results: Of the 48 M. tuberculosis that elicited MDR phenotype by the proportion and MIC methods, 2 (4.2%) isolates lacked any
of the inhA, katG and rpoBgene mutations associated with RIF and isoniazid resistance in the line probe assay method. The highest
occurring rpoB mutation conferring RIF resistance among the MDR M. tuberculosis tested was 97.3% with mutation point S531L
at a frequency of 60.4% followed by H526D mutation (22.9%), H536Y (8.3%) and D516V (6.3%). The S315T mutation of the katG
gene was responsible for 50% of isoniazid resistance among the MDR isolates. This was followed by C-15T mutation (14.6%), S94A
mutation (12.5%) and 1194A (8.3%) of inhA mutation points.
Conclusions: Different mutations of genes encoding either drug target or the enzymes involved in drug activation such as S531L,
H526D, H536Y, D516V, S315T, C-15T and S94A were detected in MDRTB isolates. The study also showed that lack of inhA, katG and
rpoB resistance defining mutations may not be sufficient as markers of susceptibility to anti-TB drugs. There is need for a nation-wide
study of the pattern of mutations of drug/enzyme gene targets in MDRTB in Nigeria.

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Keywords thereby causing a lethal effect on the M. tuberculosis; if there is a
Pulmonary tuberculosis, M. tuberculosis, Gene Mutations. mutation of the KatG gene, the drug will not be converted and the
organism appears resistant to the drug [5].
Introduction
Tremendous improvement in the diagnosis and treatment of RpoB gene is the RNA polymerase B gene responsible for the
tuberculosis had been made globally. However, pulmonary binding of Rifampicin antibiotic to the RNA of organism thereby
tuberculosis (PTB) remains one of the major causes of death in causing a lethal effect in the organism. If there is resistance, the
adults and children. According to the World Health Organisation [1], drug will not bind to the mRNA of the organism and the organism
a fewer number of people who presented for MDRTB diagnosis were appears resistant to the Rifampicin drug.
detected and Nigeria is one of the five countries in the world with
underreported cases of TB cases (including MDRTB). In countries The absence of mutations does not preclude resistance to
with underreported cases, intensified efforts are required to reduce antimycobacterial drugs because other mechanisms of resistance
underreporting and improve access to diagnosis and treatment. may be involved [8,9]. Mycobacterium tuberculosis and other
World Health Organization further emphasised some of the strategies members of the M. tuberculosis complex use several strategies to
for detecting more MDRTB and recommends detection of MDR resist the action of antimicrobial agents and these are: mycobacterial
TB using bacteriological confirmation of TB and testing for drug cell is surrounded by a specialized, highly hydrophobic cell wall
resistance using rapid molecular tests, culture methods or sequencing that results in decreased permeability to many compounds, active
technologies [1]. The M. tuberculosis strains resistant to multiple drug efflux systems and degrading or inactivating enzymes and
anti-TB drugs are becoming increasingly common. These include mutation of the genes responsible for drug targets. The genes
improvement in the detection of TB and increasing bacteriological that are associated with these functions, have since been found
confirmation of TB cases, increasing coverage of testing for in M. tuberculosis [9,10]. Considerable work has been done on
MDRTB among those bacteriologically confirmed TB cases. the characterization of drug-resistant mycobacteria. Structural
or metabolic genes encoding either the enzymes that activate
The danger of M. tuberculosis strains resistant to multiple anti-TB antimycobacterial drugs or the protein targets of drug action that
drugs was since reported as becoming increasingly common since lead to a high level of resistance to a single drug when the genes are
2003 [2]. Multidrug-resistant tuberculosis (MDR-TB) is the main altered by mutation have been identified. In most cases, multidrug-
indicator of previous treatment in tuberculosis (TB) patients and resistant isolates have accumulated independent mutations in
MDR-TB among treatment-naïve patients indicates infection with several genes [10].
drug-resistant Mycobacterium tuberculosis strains, and such cases
are considered primary drug-resistant cases [3]. The designated mutant katG in LPA is S315T. The second target
gene in the isoniazid pathway is inhA, which encodes an enoyl
Mutations associated with changes in the primary targets of ACP synthase involved in the biosynthesis of mycolic acid, an
activated INH are enzymes peroxidases involved in the biosynthesis important lipid component in the cell wall of M. tuberculosis. The
of cell wall mycolic acids have been shown to contribute to INH designated mutant inhA in LPA are C15T, A16G, T8C and T8A.
resistance [4]. Resistance has been linked to mutation of rpoB, For rifampicin, resistance is mediated by mutations in rpoB gene
kat G and inh A genes for resistance to rifampicin and isoniazid that encoded the beta subunit of M. tuberculosis RNA polymerase
respectively [5]. enzyme.

MDR M. tuberculosis strains seem to have accumulated Rationale for the study
independent mutations in genes encoding either the drug target Mutations of drug target genes in M. tuberculosis remain very
or the enzymes involved in drug activation. In a few reference important in the emergence of MDRTB. Mutation patterns in M.
laboratories in Nigeria, MDR-TB is diagnosed phenotypically tuberculosis vary from strains to strains and with geographical
by the use of the drug sensitivity testing based on proportion and environmental differences. The Line Probe Assay detects
method using LJ medium supplemented with isoniazid at 0.2 MDR-TB based on the detection of mutations in genes targeted
ug.mL concentration and rifampicin at 40 ug/mL. Although this by isoniazid and rifampicin [7]. They include katG, a catalase
method has been found to be very reliable in diagnosing MDR- peroxidase gene involved in the activation of isoniazid. The rpoB
TB, it is time consuming, laborious and lacks information on the mutations included in the LPA DNA strip technology are D516 V,
mechanism of resistance [6,7]. To improve on MDR-TB diagnostic H526Y, H526D and S531L. These markers are strongly associated
turnaround time, genotypic testing based on the use of line probe with MDR-TB in M. tuberculosis by eliciting MDR phenotype
assay (LPA) is also performed. The LPA detects MDR-TB based [10]. There is therefore the need to understand the mutation types
on the detection of mutations in genes targeted by isoniazid and and frequency in the MDR-TB among circulating strains of M.
rifampicin from the DNA sample of sonicated Mycobacterial tuberculosis in the study area, which is presently having limited
cells. They include katG, a catalase peroxidase gene involved in information regarding the most frequent mutations in the katG
the activation of isoniazid. KatG gene is for producing hydrogen and inhA pathways for isoniazid resistance and the rpoB gene for
peroxide that will convert INH from inactive form to active form rifampicin resistance.

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Objectives of the study microtitre plates with U-shaped wells as described by Leite et
This study is to (i) determine the prevalence of MDR-TB isolated al., 2000 [12]. The wells of the microtitre plates were filled with
in Lagos State, Nigeria (ii) assess the occurrence and frequency 0.1 ml amounts of Middlebrook 7H9 broth, supplemented with
of gene mutations associated with rifampicin (RIF) and isoniazid oleic acid, albumin, dextrose and, catalase (OADC) enrichment.
(INH) resistance from the isolated MDRTB, (iii) compare different The stock suspensions of drugs were diluted in Middlebrook 7H9
methods of detecting MDRTB and (iv) investigate whether all the broth and seven serial dilutions for each drug and the microtitre
M. tuberculosis which elicited MDRTB phenotype had mutation plates were stored at –25°C until use. The antibiotics gradient was
markers. 3.2-0.05 µg/ml for INH, 16-0.25 µg/ml for RIF and, 32-0.5 µg/
ml for STR and ETM. Each well was inoculated with 5 µl of 0.5
Study site McFarland standard bacterial suspension. A well without anti-TB
This study was carried out at the Nigerian Institute of Medical drugs was also inoculated with 10-2 dilution of 0.5 McFarland
Research Yaba, Lagos, Nigeria. standard as growth control. The microtitre plates were sealed,
placed in plastic bags and incubated at 37°C for 14 days in a
Study design moisturized incubator. The Minimum Inhibitory Concentration
This was a cross-sectional study conducted on stocked MDRTB (MIC) was defined as the lowest drug concentration that exhibited
Isolated from TB patients in Lagos State between May 2012 and no growth by visual reading, and the strains were considered
October 2016. susceptible for each drug if MICs were below or equal to the
critical concentration as described by Heifets and Iseman [13].
Ethical considerations The results were evaluated after 7, 10 and 14 days and they were
The study was approved by the Institutional Review Board of the compared with those obtained by the proportion method using
Nigerian Institute of Medical Research, Yaba, Lagos. the critical concentration of drugs.

Sample size After 7 days, 30 μl of 0.02 per cent resazurin sodium salt solution
A total of 306 M. tuberculosis were studied out of which 48 was added to each well and again incubated for a further 24 h at
MDRTB were confirmed by proportion method, Resazzurin 37oC. A change in colour of the resazurin dye from blue to pink
Microtitre Assay (REMA) and Line Probe Assay (LPA) for drug was considered as positive growth and MIC was determined
susceptibility and resistance testing. as corresponding to the concentration in the last blue colour in
a row. All the experiments were repeated in duplicates. Isolates
Data Analyses with MICs of INH <0.25 μg/mL and RIF ≤1 μg/mL were defined
Data obtained after questionnaire administration were double- as being sensitive to INH and RIF, respectively. Reporting
entered into Microsoft excel 2007 version and Epi INFO version recommendations have been addressed by the Clinical and
6.1. They were cleaned and validated for completeness and error Laboratory Standards Institute [11].
before export to Statistical Package for Social Science (SPSS
version 26) where analyses were done. Demographic variables DNA extraction for molecular assays
such as age, sex, education, occupation and clinical data such as DNA extraction was carried out by sonication on the cultured
the presence of fever, cough, hemoptysis, night sweat, diabetes, organisms, followed by heat killing at 80◦C for 30 minutes,
and HIV were used as covariates and summarized as frequency and followed by multiplex amplification with biotinylated primers
percentages (%) as well as mean + standard deviation (SD). Chi - and reverse hybridization. Identification of MTBC and NTM
square(χ2)and Fisher Exact (when expected frequency (e) < 5) test species were carried out by using specific sets of primers
was used to evaluate the relationship between proportion method designed to amplify a species-specific 23S rRNA gene sequence
of DST and LPA as diagnostic tools for MDR TB infections. of Mycobacterium species. One ml of MGIT 960 culture of
reference or clinical M. tuberculosis isolate was heated with 40 mg
Drug Susceptibility Testing (DST) Chelex-100 (Sigma-Aldrich, St. Louis, MO, USA) at 95°C for 20
Inoculum preparation - Freshly grown colonies (> 50) of M. min and then centrifuged at 12,000 × g for 15 min. For a PCR, 2 μl
tuberculosis isolates from LJ medium were transferred to a tube of supernatant was used as a source of DNA.
containing 3-4 ml phosphate-buffered saline and 6 to 9 sterile
glass beads. Tubes were vigorously agitated on a vortex mixer This technique involved DNA amplification targeting the 23s
and clumps were allowed to settle for 30 min. The suspensions rRNA region of NTM isolates, followed by reverse hybridisation
were transferred to sterile tubes and adjusted with phosphate buffer to specific oligonucleotide probe immobilised on membrane strips.
saline to equal the density of 0.5 McFarland standard for use as the The kit for common Mycobacteria (CM), which identifies 15
standard inoculum in the Broth microdilution method (BMM) and Mycobacterium species, including M. tuberculosis complex, was
reazurin microtitre method (REMA) [11]. used [14].

REMA: Resazzurin Microtitre Assay (REMA) technique was Amplification mixture (45 μl) was prepared in DNA free room,
performed using Middlebrook 7H9- liquid medium in 96-well including 5 μl extracted DNA (20-100 ng DNA) in the reaction

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mixture contained 35 μl primer nucleotide mix, 5 μl 10 × and 2 min at 58°C, followed by 20 additional cycles of 25
polymerase incubation buffer for HotStarTaq (QIAGEN, Hilden, s at 95°C, 40 s at 53°C, and 40 s at 70°C, ending with a final
Germany), 2 μl 25 mM MgCl2 solution, 0.2 μl Hot StarTaq and 3 extension step of 8 min at 70°C. Hybridization and detection
μl water (biology grade water). Amplification was carried out in were performed with a TwinCubator semi-automated washing
a thermal cycler (MJ Research, PTC-100 Thermal Cycler, GMI, and shaking device according to the manufacturer's instructions
Inc, USA), which involved 01 cycles of denaturation solution and using the reagents provided with the kit. Briefly, 20 μl of
(DEN) at 95°C for 15 min, annealing of primers at 95°C for 30s, denaturation solution was mixed to 20 μl of amplified sample
2 min at 58°C for 10 cycles, then 20 cycles at 95°C for 25 s, 53°C and incubated at room temperature for 5 min. One milliliter of
for 40 s and 70°C for 40 s and final primer extension at 70°C, 8 prewarmed hybridization buffer was added before the membrane
min for 01 cycle. The amplified products were stored at +8 to strips were placed and shaken in the hybridization solution for 30
−20°C until hybridization was done in a hybridization machine min at 45°C. After two washing steps, a colorimetric detection
(Profiblot; Tekan, Maennedorf, Switzerland). The hybridization of the hybridized amplicons was obtained by the addition of
included the chemical denaturation of the amplification products, the streptavidin alkaline phosphatase conjugate. All LPA runs
hybridization of a single-stranded, biotin-labelled amplicon to adhered to ISO 15189 standards, which require the use of an
membrane-bound probes, stringent washing, the addition of a ATCC M. tuberculosis H37Rv laboratory strain for positive
streptavidin/alkaline phosphatase conjugate, and an alkaline control. Two negative controls were used to test for area-specific
phosphatase mediated staining reaction. Here, 20 μl of DEN contamination [15].
(blue) was dispensed in the corner of each well and then 20
μl of amplified product was added and incubated for 5 min at Results
room temperature. Then 1 ml of pre-warmed hybridization Of the 306 M. tuberculosis isolates, 48 MDR phenotype by the
buffer (HYB, Green) was added, followed by gentle shaking proportion method showing a prevalence of 15.7%. Tables 2
until a homogenous colour was developed. Now strip was placed and 3 showed four (4) distinct resistance patterns were observed
in a manner to make sure complete flooding of solution over among the MDR isolates with INH RIF occurring the most
strips. Then tray was placed in TwinCubator and was incubated (48/306) and INH RIF EMB occurring the least (2/306). Table
for 30 min at 45°C, followed by complete aspiration of HYB. 2 showed different resistance types which were polyresistance
Washing was done by 1 ml of stringent wash solution to each in 30/306 isolates and monoresistance for INH (16/306) and RIF
strip and incubated for 15 min at 45°C in TwinCubator. Again, (4/306). On the whole, 208 (68%) isolates were found to be pan
strips were washed once with 1 ml of ringer solution for 1 min susceptible. Of the 48 MDR M. tuberculosis isolated tested, two
on TwinCubator. Then 1 ml of diluted conjugate was added to (2) isolates did not harbour any of the four (4) rpoB mutations
each strip and incubated for 30 min on TwinCubator. Strips were associated with RIF resistance and lack of mutations associated
washed again with 1 ml of ringer solution for 1 min, after that 1 with INH in katG or inhA genes were detected in nine (9)
ml of diluted substrate were added to each strip and incubated isolates. Table 3 showed that the highest occurring rpoB mutation
for 3-20 min in the absence of light without shaking. Rinsing conferring RIF resistance among the 48 MDR M. tuberculosis
was done twice with distilled water to stop the reaction. Strips tested was 97.3% with mutation point S531L at a frequency of
were removed and dried between two layers of absorbent paper. 60.4% followed by H526D mutation (22.9%), H536Y (8.3%) and
Evaluation and interpretation of results were done based on the D516V (6.3%). The S315T mutation of the katG gene (Table 3)
presence and absence of different bands and compared with was responsible for 50% of isoniazid resistance among the MDR
reference band as provided in the kit [14]. isolates. This was followed by C-15T mutation (14.6%), S94A
mutation (12.5%) and 1194A (8.3%) of inhA mutation points.
Standard strain of M. tuberculosis complex H37 Rv, Mycobacterium
fortuitum, Mycobacterium intracellulare and M. abscessus Table 4 showed the pattern of the detection of MDRTB Isolates
obtained from National TB Reference Laboratory, Nigerian by MIC, Proportion and LPA methods. Of the 48 MDRTB isolates
detected by proportion method, 95.8% and 81.25% were detected
Institute of Medical Research, Yaba, Lagos were used as control
by MIC and LPA methods, respectively.
in this study.
Table 1: Age, Gender and Treatment History of the Participants with
Genotype MTBDR assay
MDRTB Isolates.
The Genotype MTBDR assay (Hain Lifescience, Nehren,
Germany) was performed as recommended by the manufacturer. Gender Treatment History
Age No with
The amplification mixture contained 35 μl of primer- distribution MDRTB New
Male Female Retreatment
nucleotide mix provided in the kit, 5 μl of 10× Taq polymerase treatment
incubation buffer containing 2 mM of MgCl2, 1 to 2 unit(s) 18-35 22 2 19 3 (37.5) 2 (25)
of thermostable Taq DNA polymerase, and 5 μl of extracted ≥ 36 26 27 3 6 (54.5) 5 (45.5)
chromosomal DNA solution in a final volume of 50 μl. The
following amplification parameters were used: 5 min of Mean=34.3 years
denaturation at 95°C, followed by 10 cycles of 30 s at 95°C

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Table 2: Susceptibility Pattern of the M. Tuberculosis Isolated from the et al., [7] where only 95 % detection of resistance in MDRTB was
Study Participants. detected using LPA method. This study showed that phenotypic
No of M. methods using proportion and /or MIC methods appeared to be
Resistance Types %
Tuberculosis better options for detection of MDRTB and may be required for all
Mono Resistance 20 6.5 LPA negative MDRTB test.
(INH OR RIF)
The designated mutant of katG in LPA was S315T while the
INH and RIF Resistance (MDRTB) 48 15.7
designated mutant of inhA in LPA are C15T, A16G, T8C and T8A
Polyresistance (INH, RIF, EMB) 0 0.0 while that of rpoB of is S531L. The results of this study showed
PAN Susceptible 238 77.8 that 50% and 60.4% of the isolated MDRTB had S315T and S531L
TOTAL 306 100 mutants of katG and rpoB respectively while 14.6% C15T mutants
of inhA responsible for INH resistance (Table 3). For rifampicin,
Table 3: Occurrence and Frequency of rpoB, Kat G and InhA Mutation resistance is mediated by mutations in rpoB gene encoded the beta
Pattern in the MDR M. Tuberculosis. subunit of M. tuberculosis RNA polymerase enzyme.
Drug Mutation No of MDRTB Frequency
Target Point Showing the Mutation (N= 48) (%) Occurrence and frequency of the different mutants in different
S531L 29 60.4 environments may cause strains to strains variations and may in
turn, influence the diagnostic value of the LPA method.
H526D 11 22.9
rpoB
H536Y 4 8.3 Table 4 showed the percentage detection of MDRTB with minimum
D516V 3 6.3 inhibitory concentration (MIC), Proportion and LPA methods (p =
S315T 24 50.0 0.01). This showed that there was significant difference in the results
kat G for MDRTB recorded when minimum inhibitory concentration
S315N 3 6.3
(MIC), proportion and LPA methods were used. Nine (18.75%)
S94A 6 12.5
of the isolates were false-negative by LPA which could be due
C-15T 7 14.6 lack of inhA, katG and rpoB resistance defining mutations This
inhA
1194A 4 8.3 difference implied that drug resistance could be due to combined
121TN 1.0 2.1 effect of mutation and other mechanisms of drug resistance or it
could be due to the absence of the mutation points among MDRTB
Table 4: Detection of MDRTB Isolates by Minimum Inhibitory circulating in Lagos. Louw et al., [5]; Lana et al.,[7] and Raheem
Concentration (MIC), Proportion and LPA methods. N= 48. et al.,[9] had earlier suggested that other biological mechanisms
Method Susceptible Percentage (%) P Value such as efflux pumps could be a factor that could be responsible
for resistance even where mutation of the rpoB gene and other
MIC 46 95.8 0.01
drug target genes did not occur in the M. tuberculosis. Variations
Proportion 48 100 of mutations by regions and environment could be responsible for
LPA 39 81.25 failure to detect MDRTB by LPA in addition to other factors.

P = 0.01. This showed that there was significant difference in Conclusions

the results for MDRTB recorded when minimum inhibitory Though 84.3% of the TB isolates were drug susceptible, 15.7% of
concentration (MIC), proportion and LPA methods were used. the isolates were Multidrug Resistant TB (MDRTB) in this study.
Different gene mutation markers such as inhA, katG, rpoB S531L,
Results and Discussions H526D, H536Y, D516V, S315T, C-15T and S94A were detected in
Table 2 showed the prevalence of the resistant or susceptibility the MDRTB isolates. 81.25% of the MDRTB were detected by line
patterns of all the M. tuberculosis isolates with 6.5%, 15.7% and probe assay indicating rpoB, katG and inhA mutations, which are
77.8% mono resistance, MDR and pan susceptibility respectively. determinants of resistance to Rifampicin and Isoniazid. However,
Mutations associated with changes in the primary INH (kat G and 18.75% of the resistance detected by proportion method (which
InhA) and Rifampicin (rpoB) drug targets were detected in this indicated 100% of the MDRTB) were not detectable by LPA
study. The MDRTB prevalence of 15.7 % recorded in this study method. MDRTB were detected more with phenotypic (proportion
was similar to the study of Lana et al., [7] in a study carried out method) than those based on drug target gene mutations alone. The
in Lagos. However, INH MDRTB detected by LPA in this study study showed that the absence of inhA, katG and rpoB resistance
was 39 out of 48 (81.25%) unlike the 48/48 (100%) by proportion defining mutations may not be enough as markers of susceptibility
method. Compared to the proportion method, the MIC and LPA to anti-TB drugs. Phenotypic methods using proportion and or
methods of drug susceptibility testing correctly diagnosed 95.8% MIC methods should be performed on all LPA negative MDRTB
and 81.25% cases, respectively. The study, therefore, showed samples to reduce false-negative results and increase detection of
discordance in resistant results between LPA and Proportion MDRTB cases.
methods. This finding is similar to the study conducted by Lana
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References 9. Raheem Toyosi Yekeen, Iwalokun Bamidele, Fowora Muinah,
1. Global tuberculosis report. World Health Organization. 2020. et al. Are Drug Efflux Genes Present Among Mycobacterium
2. Espinal MA. Time to abandon the standard retreatment tuberculosis Isolates from Patients in Lagos, Nigeria?. Journal
regimen with first-line drugs for failures of standard treatment. of Biosciences and Medicines. 2020; 8: 86-98.
Int J Tuberc Lung Dis. 2003; 7: 607-608. 10. Somoskovi Akos, Linda M Parsons, Max Salfinger. The
3. Da Silva Garrido M, Ramasawmy R, Perez-Porcuna TM, et molecular basis of resistance to isoniazid, rifampin, and
al. Primary drug resistance among pulmonary treatment-naïve pyrazinamide in Mycobacterium tuberculosis. Respir Res.
tuberculosis patients in Amazonas State, Brazil. Int J Tuberc 2001; 2: 164-168.
Lung Dis. 2014; 18: 559-565. 11. Gail L. Woods, Barbara A. Brown-Elliott, Patricia S, et
4. Slayden RA, Barry CE III. The genetics and biochemistry of al. Susceptibility Testing of Mycobacteria, Nocardia spp.,
isoniazid resistance in Mycobacterium tuberculosis. Microbes and Other Aerobic Actinomycetes. Clinical and Laboratory
Infection. 2000; 2: 659-669. Standards Institute. 2018.
5. Louw Gail E, Robin M. Warren, Nicholaas C. Gey van 12. Leite CQF, Beretta ALRZ, Anno IS, et al. Standartization of
Pittus, et al. Rifampicin Reduces Susceptibility to ofloxacin broth microdilution method forMycobacterium tuberculosis.
in Rifampicin-Resistant Mycobacterium tuberculosis through Mem Inst Oswaldo Cruz. 2000; 95: 127-129.
Efflux. Am J Respir Crit Care Med. 2011; 184: 269-276. 13. Heifets LB, Iseman MD. Individualized therapy versus
6. Maurya AK, Nag VL, Kant S, et al. Evaluation of an standard regimens in the treatment of Mycobacterium avium
immunochromatographic test for discrimination between infections. Am Rev Respir Dis. 1991; 144: 1-2.
Mycobacterium tuberculosis complex & non tuberculous 14. Lee H, Yoon TJ, Weissleder R. Ultra-sensitive detection of
mycobacteria in clinical isolates from extra-pulmonary bacteria using corel-shell nano-particulas and an NMR – filter
tuberculosis. Indian J Med Res. 2012; 135: 901-906. system. Angew Chem Int Ed Engl. 2009; 48: 5657-5660.
7. Lana Dinic, Patrick Akande, Emmanuel Oni Idigbe, et al. 15. GenoType MTBDRplus manual: In Molecular genetic assay
Genetic Determinants of Drug-Resistant Tuberculosis among for identification of resistance to rifampicin and/or isoniazid of
HIV-Infected Patients in Nigeria. J Clin Microbiol. 2012; 50: the Mycobacterium tuberculosis complex. Hain Lifescience.
2905-2909. 2009; 23-42.
8. Barbara A. Brown-Elliott, Gail L. Woods. Antimycobacterial
Susceptibility Testing of Nontuberculous Mycobacteria. J
Clin Microbiol. 2019; 57: e00834-e00819.

© 2024 Raheem TY, et al. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License

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