Research Article ISSN 2639-9458

Microbiology & Infectious Diseases

Phenotypic Detection of Rifampicin and Isoniazid Resistance Pattern of

Mycobacterium Tuberculosis Auramine Positive Isolates in Mtata, South Africa
Osuji C Emmanuel1* and Okezie C Okamgba2

1
BobManuel Path Care, Johannesburg, South Africa. *
Correspondence:
Osuji C Emmanuel, BobManuel PathCare, Johannesburg, South
2
Department of Medical laboratory Science, Faculty of Medicine Africa.
and Health Sciences, Abia State University, Uturu, Abia State,
Nigeria. Received: 02 Feb 2022; Accepted: 07 Mar 2022; Published: 11 Mar 2022

Citation: Emmanuel OC, Okamgba OC. Phenotypic Detection of Rifampicin and Isoniazid Resistance Pattern of Mycobacterium
Tuberculosis Auramine Positive Isolates in Mtata, South Africa. Microbiol Infect Dis. 2022; 6(2): 1-4.

ABSTRACT
Background: The increase prevalence of Isoniazid (INH) and Rifampicin (RIF) resistant Tuberculosis (TB) has
become a significant challenge for TB control for over 10 years.

Aim: The study assessed the phenotypic detection of rifampicin (RIF) and isoniazid (INH) resistant pattern of
Mycobacteria tuberculosis among patients attending hospitals in Mtata, South Africa.

Methodology: Three hundred and fifty (350) patients age between 10-80 years attending Nelson Mandela Hospital
Mtata were recruited for the study. Ethical approval was obtained from the Ethics Committee of Nelson Mandela
Hospital Mtata. Sputum specimens were investigated employing Auramin fluorescence microscopy technique.
Drug susceptibility testing (DST) was carried out using cultural technique and conventional drugs susceptibility
testing method.

Results: Result showed that 28.6% of the isolates were positive on culture. For INH testing, 27% were sensitive,
68% were resistant where as 5% had lost viability to INH. For RIF, 28% were sensitive, 69% were resistant and
3% had lost viability to RIF. The rural settlers has 55% INH resistance and 26% RIF resistance while in urban
area 35% and 25% were resistance to RIF and INH respectively.

Conclusion: Result from this study showed that INH and RIF resistance were very high among the patients
attending Nelson Mandela Hospital and other treatment centers in Mtata, South Africa.

Keywords from all parts of the world, however, over 95% of cases and deaths
Drug sensitivity testing (DST), Micobacterium tuberculosis occur in developing countries [4].
(MBT), Auramin, Isoniazid (INH), Rifampicin (RIF).
Multidrug Resistance – Tuberculosis (MDR- TB) rate of 2.6%
Introduction among new cases and 17.6% among retreatment cases was reported
Mycobacterium tuberculosis (MBT) infection remains a common at the latest National Survey in Nepal [5]. The emergence of MDR-
infectious disease worldwide, with up to 10 million reported cases TB is widely considered to be a serious threat to global TB control,
and 2 million deaths per year [1]. Meanwhile, the incidence of and in Nepal, being geographically situated between China and
this disease seemed to have declined during the 1970s and early India which carry almost 50% of worlds MDR- TB burden, drug
1980s because of the success in chemotherapy. However, there has resistance TB is emerging as a national problem [6-8]. Isoniazid
been a global resurgence of tuberculosis since the late 1980s [2]. (INH) is one of the most effective agents for the treatments of
In Hong Kong, 6607 cases were recorded in the year 2002, at the tuberculosis and is the backbone of the modern short-course. As
rate of 97/100 000 people [3]. Tuberculosis (TB) has been reported one of the first-line anti –TB drugs, INH has been used in TB
Microbiol Infect Dis, 2022 Volume 6 | Issue 2 | 1 of 4
control programs for greater than 50 years and for the treatment of Results
latent Mycobacterium tuberculosis (LMTB) Infection to prevent Table 1: Total Sample Treated With %AUR Positive.
the active disease and subsequent TB transmission. However, the AUR Total %
Total sample
rapid increase of INH resistance in recent years poses a serious +ve AUR +ve
threat to global TB control, especially in China. WHO report 2008, 350 100 28. 6
estimated that the INH resistance rate in China is 41.2%, which is
Table 2: Various Ages of the Patients Sampled.
much higher than the global estimates of 13.3% [9]. Considering
AUR % AUR
the 13.1% estimated INH resistance rate in 2000 reported by Age Samples
+ve AUR +ve -ve
National survey [10]. The dissemination of INH resistance TB 10-20 50 5 5 45
in china is dramatically high in the last 10 years. The speedy 21-30 50 10 10 40
dissemination of INH resistance is of public health concern as it 31-40 50 13 13 37
could lead to increased prevalence multidrug resistant TB (MDR- 41-50 50 15 15 35
TB). MDR-TB is defined as TB resistance to at least INH and RIF 51-60 50 15 15 35
in some instances to as many as seven anti-TB drugs [11]. By now, 61-70 50 20 20 30
China has the largest number of MDR- TB cases in the world, 71-80 50 22 22 28
and many extensively drug- resistant TB cases have been reported Total 350 100
[12,13]. Recent increases both in INH- resistant and multi drug
–resistant (MDR) tuberculosis have been jeopardizing the effects Table 3: Rate of Positive MBT among Rural and Urban Settler.
of global TB control through the implementation of the directly Location
Sample from AUR % AUR
observed treatment short course (DOTS) [14,15]. location +ve AUR +ve -ve
Rural Settlers
175 70 70 105
Ages 10-80
In China, DOTS program was adopted since early 1990s, but the Urban Settlers
increased prevalence of drug resistant TB has become a significant 175 30 30 145
Ages 10-80
challenge for TB control in last ten years. The prevalence of MDR- 350 100 100
TB and INH resistant was estimated at 8.3% and 41.2 % respectively
among all cases in china which were both high compared to the Table 4: Indicates Percentage Resistance and Sensitive of INH
global estimates of 4.8% and 13.3% [16]. The development of Respectively.
INH resistance is a common first step in the evolution to MDR Ages INH
INH INH % %
DOT LV
[17]. Thus, there has been considerable interest in identifying S R INH R INH S
the molecular basis of INH resistance and understanding the 10-20 5 4 1 + -
21-30 10 5 4 + 1
transmission pattern of the INH resistant MTB strain.
31-40 13 2 9 68 27 - 2
41-50 15 2 13 - -
Materials and Method 51-60 15 2 12 - 1
This study was conducted in TB department Nelson Mandela 61-70 20 4 16 - -
Hospital, Mtata, South Africa. A 350-sputum specimen were 71-80 22 8 13 + 1
obtained from patients on DOTS and patients on treatment among 100 27 68
rural and urban settlers.
Table 5: Percentage Resistance and Sensitive of RIF Respectively.
Methods Ages RIF
RIF RIF % %
DOT LV
Specimen were obtained in a sterile, leak proof, wide mouth S R RIF R RIF S
10-20 5 3 2 + -
transparent and stoppered plastic containers. Sputum smears
21-30 10 4 6 + -
were made on slides, stained using Auramine, and subsequently
31-40 13 5 8 69 28 - -
viewed using fluorescence microscopy. All acid fast positive
41-50 15 4 11 -
specimen were cultured on Lowenstein –Jensen (LJ) medium. 51-60 15 3 10 - 2
Culture positive specimen were assessed for drug resistance using 61-70 20 4 16 - -
phenotypic conventional DST. Proportion method was used on LJ 71-80 22 5 16 1
medium for DST at critical concentration of 0.2 ug/ml for INH 100 28 69
and 40 ug/ ml for RIF. The bacterial culture plates were read after
6 weeks of incubation at 37°C and the strains were considered Table 6: Percentage Resistance of INH among Rural Settlers.
resistant if the proportion of resistant bacteria was higher than 1%. Aur % % %
Location Sample INHR INHS LV DOT
+ve Aur INHR INHS
Statistical Methods Rural
Chi square was used for analysis of values. Level of significance Settlers 175 70 70 35 30 35 30 5 -
was set at p < 0.05. The statistical analysis was done using SPSS Ages
statistical software package (version 21). 10-80

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Table 7: Percentage Resistance of RIF among Rural Settlers. finding in (table 5) revealed that RIF had the highest resistance
Location Sample
Aur %
RIFR RIFS
% %
LV DOT
(69%) followed by INH (68%). This is difference from the
+ve Aur RIFR RIFS findings of Annelies et al. [25] reported that resistance was 93%,
Rural 56%, 41%, and 18% to INH, RIF, STR and EMB respectively and
Settlers 175 30 30 26 4 26 4 - -
a lower prevalence of resistance was reported by Rivera et al. [26]
Ages
who studied 188 MTB isolate in Philippines and noted that drug
10-80
resistance represented 17, 6% of isolates and in Qatar, by Al –
Table 8: Percentage Resistance of RIF among Urban Settlers. Marri [27] who found 6 (15%) among 406 MTB isolate. It was
Aur % % discovered that rural settlers are more affected with TB compare
Location Sample RIFR RIFS %RIFS LV DOT
+ve Aur RIFR to urban settlers with a higher multidrug resistance. The rural
Urban settlers has 55% INH resistance and 26% RIF resistance while in
Settlers 175 55 55 24 27 24 27 4 + urban area there is less percentage of 35% and 25% of RIF and
Ages INH respectively. This may be non-implementation of directly
10-80 observed therapy short course (DOTS) among the rural settlers.
In my study, the incidence of drug resistant is higher compared
Table 9: Percentage Resistance of INH among Urban Settlers.
to that reported in a previous study conducted by Massi et al. [28]
Aur % % %
Location Sample
+ve Aur
INHR INHS
INHR INHS
LV DOT in Makassar, Indonesia, where the proportion of Mycobacterium
Urban tuberculosis resistant to at least one first – line TB drug reached
Settlers 175 45 45 25 18 25 18 2 + 40% in new and retreatment cases. The proportion of DR in my
Ages study is also higher than that reported Nana et al. [29] in China
10-80 where 33.3% isolate were resistant to all first line TB drugs. The
List of Abbreviations: finding in tables 6 and 7 of the rural settlers have higher percentage
MTB: Mycobacterium Tuberculosis; INHR: Isoniazid Resistant; RIF: of INHR and RIFR with 35% and 26% respectively compared to
Rifampicin; INH: Isoniazid; RIFR= Rifampicin Resistant; MDR: table 8 and 9 of the urban settlers with percentage of RIFR and
Multidrug – Resistant; AUR: Auramin; DOTS: Directly Observed INHR 24% and 25% respectively , this is attributed to the rural
Treatment Short Course; LV: Lost Viability; DST: Drug Susceptibility settlers not observing or making use of DOT. It was discovered
testing. that rural settlers are more affected with TB compare to urban
settlers with a higher multidrug resistance.
Discussion
Tuberculosis, one of the oldest recorded human diseases is still Conclusion
one of the biggest killers among the infectious diseases ravaging Result from this study showed that INH and RIF resistance were
the globe despite the use of a live attenuated vaccine and several very high among the patients attending Nelson Mandela Hospital
antibiotics. The development of drug resistance in the population and other treatment centers in Mtata also rural settlers are more
has increased the possibility that Tuberculosis may once again affected with a high multidrug resistance than urban settlers in
become an incurable disease [18]. The emergence and spread South Africa.
of DR- TB threatens TB control and has become a major Public
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© 2022 Emmanuel OC & Okezie CO. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License

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