Prevalence Of Drug-resistant Strains Research Articles

Factors associated with tuberculosis drug resistance among presumptive multidrug resistance tuberculosis patients identified in a DRTB surveillance study in western Kenya

Multidrug-resistant tuberculosis (MDR-TB) is caused by M. tuberculosis (Mtb) with resistance to the first-line anti-TB medicines isoniazid (INH) and rifampicin (RIF). In Western Kenya, there is reported low prevalence of drug resistant strains among HIV tuberculosis patients, creating a need to determine factors associated with drug resistance patterns among presumptive MDR-TB patients. To determine factors associated with drug resistance patterns among presumptive MDR-TB patients in western Kenya. Three hundred and ninety (390) sputum sample isolates from among presumptive multidrug TB patients, were analyzed for TB drug resistance as per Ministry of Health (MoH) TB program diagnostic algorithm. Frequency and percentages were used to summarize categorical data while median and interquartile range (IQR) were used for continuous data. Multivariable logistic regression was carried out to identify factors associated with TB drug resistance. Out of 390 participants enrolled, 302/390 (77.4 %) were males, with a median age of 34 years. The HIV-infected were 118/390 (30.3 %). Samples included 322 (82.6 %) from presumptive patients, while 68/390 (17.4 %) were either lost to follow-up patients, failures to first-line treatment or newly diagnosed cases. A total of 64/390 (16.4 %) of the isolates had at least some form of drug resistance. Out of 390, 14/390 (3.6 %) had MDR, 12 (3.1 %) were RIF mono-resistance, 34 (8.7 %) had INH, while 4 (1 %) had ethambutol resistance. The category of previously treated patients (those who received or are currently on TB treatment) had a 70 % reduced likelihood of resistance (aOR: 0.30; 95 % CI: 0.13–0.70). In contrast, older age was associated with an increased likelihood of resistance to INH and RIF, with an adjusted odds ratio of 1.04 per year (95 % CI: 1.00–1.08). Prompt MDR-TB diagnosis is essential for appropriate patient care, management, and disease prevention and control. We recommend active surveillance on drug resistant TB in these regions to detect drug resistance patterns for rapid disease management.

Standardisation of an In-house Multiplex PCR Assay for Identifying Diffusely Adherent E. coli and ESBL Producing Resistant Isolates

We aimed to develop a multiplex PCR (mPCR) assay utilizing the afaC gene for the detection of DAEC pathotypes and the blaTEM gene for the identification of ESBL production in clinical E. coli isolates along with an Internal Amplification Control (IAC) to rule out false negative test results. Monoplex PCR assays were first established for the afaC gene and blaTEM gene using 60 characterized E. coli isolates from various clinical samples. Subsequently, an mPCR assay was designed to detect both the genes simultaneously along with an IAC to rule out false negative reactions. The effectiveness of this assay was validated using 80 additional clinical isolates. The overall occurrence of DAEC in the study was found to be 0.7% (1/140). ESBL production was detected in 40.7% of the tested isolates, indicating a concerning prevalence of drug-resistant strains. This study emphasizes the value of an in-house mPCR assay as a crucial tool for simultaneously identifying DAEC and ESBL E. coli strains. The inclusion of an IAC in the PCR protocol bolstered the assay’s reliability. This innovation offers a vital resource for effective infection management and contributes to the comprehension of pathogenicity and resistance mechanisms in clinical E. coli isolates.

Problems of anti-tuberculosis therapy in patients on hemodialysis

The objective: to determine possibility of prescribing and safety of various anti-tuberculosis drugs and their combinations as part of a therapeutic or preventive course of anti-tuberculosis therapy (ATBT) in patients with end-stage chronic kidney disease (CKD) receiving hemodialysis replacement therapy in the settings with high prevalence of drug resistant strains of Mycobacterium tuberculosis.Subjects and Methods. 42 patients receiving hemodialysis replacement therapy were followed-up by Central Tuberculosis Research Institute: of them, 31 (73.8%) had active tuberculosis and 11 (26.2%) had residual post-tuberculous changes in the respiratory organs. During the follow-up, the nature and severity of adverse reactions to anti-tuberculosis drugs were assessed.Results. In the patients with end-stage CKD suffering from respiratory tuberculosis, drug resistant strains prevailed – 68.2%. Some of adverse events developed when taking anti-tuberculosis drugs were as follows: deterioration of uremia in 20.5% of cases, cardiovascular complications – in 17.9%, and neurotoxic reactions – in 15.4%. Progression of uremia occurred more often in the followed-up patients taking anti-tuberculosis drugs versus hepatotoxic reactions (p = 0.02). When treating tuberculosis patients with end-stage CKD, isoniazid was discontinued due to adverse events in 12.8% of cases and pyrazinamide in 12.8% of cases, other drugs were discontinued less frequently. Compilation of individual regimens of anti-tuberculosis drugs with the consideration of drug susceptibility testing results, contraindications, and concomitant diseases made it possible to provide an adequate anti-tuberculosis therapy for patients with end-stage CKD with tuberculosis infection in 92.9% of cases for a period of more than 6 months.

Shikimate Kinase; Searching for Potential Novel Anti‐Tubercular Agents

Tuberculosis is a respiratory infection with over 10 million reported cases each year. This infection rate is responsible for over two million deaths annually, second only to HIV in fatalities among infectious diseases annually. Studies in the last decade have shown that tuberculosis rates have gradually increased. This observation suggests that there is an alarming increase in the prevalence of drug resistant strains of tuberculosis, thus the need for the discovery of novel antitubercular agents. The shikimate pathway is a seven step metabolic route that produces aromatic amino acids and other cellular metabolites. The target enzyme in this project, Mycobacterium tuberculosis Shikimate Kinase (MtSK), catalyzes the 5th step of this pathway. MtSK converts shikimate to shikimate 3 phosphate. The overall goal of this project is to express and characterize MtSK to screen for potential antitubercular agents. Initial methods included a bacterial transformation of XL1 blue competent E.coli cells. This preliminary transformation was performed using a pET‐21b plasmid with an aroK gene inserted into the multiple cloning site. Successfully transformed XL1 blue cells were cloned and a second transformation of BL21 DE3 competent cells was performed. Initial small scale expression showed the presence of a band around 20 kDa. The theoretical mass of the enzyme is 19.6 kDa which suggests MtSK was successfully expressed. Expression analysis for large scale supported data from small scale as a band of 20 kDa was present. The purity and molecular weight of MtSK were confirmed by nickel affinity chromatography as well as ESI‐LC‐MS. ATP‐dependent kinetic parameters determined via a spectrophotometric coupled assay showed a kcat of 50 ± 6 mM, KM of 0.2 ± 0.04 mM. shikimate dependent kinetic parameters showed a kcat of 55 ± 2 mM, KM of 1.8 ± 0.1 mM. Quenching intrinsic tyrosine fluorescence showed binding affinities similar to ATP and shikimate‐dependent parameters. The marine compound, avarone showed substantial quenching of intrinsic fluorescence. The inhibitory mechanism of avarone on MtSK is currently under investigation.

Shigellosis and Development of Multiple Antimicrobial Resistance Mechanisms of Shigella spp.

Shigellosis is a serious public health issue. Millions of people suffer from this deadly food and water borne disease each year. The main manifestations of affected persons are bloody diarrhea with excessive dehydration. The causative agent of this disease is the bacteria Shigella spp. which has four serogroups. Though Shigella flexneri and Shigella dysenteriae are the dominant serogroups in developing countries, reports of other serogroups, namely Shigella boydii and Shigella sonnei, in the food contaminations are available. There are seasonal variations of Shigella infection throughout the world. In Asian subcontinent, monsoon and post monsoon times are the ideal for infection. The transmission of the bacteria in human is usually caused by feco-oral route or by contaminated food and water. There are several groups of antibiotics like foscomycin, macrolide, amiglycoside, tetracycline etc. which were used before. But they are now become useless as Shigella spp. is getting resistant against those drugs. The quinolone groups of antibiotics like ciprofloxacin, ofloxacin, norflxacin, ceftriaxone etc. are the important drugs for the cure of the disease shigellosis but prevalence of drug resistant strains of Shigella spp. against those drugs are a great concern nowadays. The occurrence of plasmid mediated quinolone resistance genes (PMQR), efflux pump proteins and effective mutations at drug binding region of gyrA etc. are the major mechanisms for the development of drug resistance.

Distribution patterns of drug resistance Mycobacterium tuberculosis among HIV negative and positive tuberculosis patients in Western Kenya

Distribution patterns of drug resistance Mycobacterium tuberculosis among HIV negative and positive tuberculosis patients in Western Kenya

The Iron Chelator Desferrioxamine Increases the Efficacy of Bedaquiline in Primary Human Macrophages Infected with BCG.

For over 50 years, patients with drug-sensitive and drug-resistant tuberculosis have undergone long, arduous, and complex treatment processes with several antimicrobials. With the prevalence of drug-resistant strains on the rise and new therapies for tuberculosis urgently required, we assessed whether manipulating iron levels in macrophages infected with mycobacteria offered some insight into improving current antimicrobials that are used to treat drug-resistant tuberculosis. We investigated if the iron chelator, desferrioxamine, can support the function of human macrophages treated with an array of second-line antimicrobials, including moxifloxacin, bedaquiline, amikacin, clofazimine, linezolid and cycloserine. Primary human monocyte-derived macrophages were infected with Bacillus Calmette-Guérin (BCG), which is pyrazinamide-resistant, and concomitantly treated for 5 days with desferrioxamine in combination with each one of the second-line tuberculosis antimicrobials. Our data indicate that desferrioxamine used as an adjunctive treatment to bedaquiline significantly reduced the bacterial load in human macrophages infected with BCG. Our findings also reveal a link between enhanced bactericidal activity and increases in specific cytokines, as the addition of desferrioxamine increased levels of IFN-γ, IL-6, and IL-1β in BCG-infected human monocyte-derived macrophages (hMDMs) treated with bedaquiline. These results provide insight, and an in vitro proof-of-concept, that iron chelators may prove an effective adjunctive therapy in combination with current tuberculosis antimicrobials.

Genotyping and Drug Susceptibility Patterns of M. Tuberculosis Isolated from HIV Seropositive Patients in Tehran Iran

to investigate the prevalence and drug-resistance M. tuberculosis isolated from HIV seropositive individuals in Tehran, Iran. Human immunodeficiency virus (HIV) is one of the most important risk factors for developing active tuberculosis (TB). To determine the rate of transmission and drug resistant M. tuberculosis (MTB) strains isolated from HIV seropositive patients in Tehran province, Iran. This study consecutively enrolled 217 TB/HIV coinfected patients from April 2018 to August 2019 at Emam Khomeini referral hospital and 5 other health centers in Tehran province. The isolates were genotyped using 15 loci Mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR). Minimum inhibitory concentration (MIC) was determined for 6 drugs. In addition, mutations were assessed in rpoB, katG, inhA, and ahpC genes using Reverse Blot Hybridization Assay System. A 20 (9.2%) patients were culture-positive for M. tuberculosis and typed by MIRU-VNTR, 13 (65%) strains formed 5 clusters, but 6 (30%) isolates had a unique pattern. The total Hunter-Gaston discrimination index (HGDI) for all 15 loci was 0.846, and the cluster size was 2 to 4 patients. The estimated proportion of recent transmission was 45%. The mutation was identified in 1 isolate, lost inhAW1 and mutation in MT1 loci, which was resistant to isoniazid (INH). Moreover, 1 (5%) and 3 (15%) isolates were resistant to INH and ethambutol (EMB), respectively, of which 1 was resistant to INH and EMB. The transmission rate of TB in HIV patients was relatively high; however, the prevalence of drug-resistant strains and TB infection in females was insignificant in this study (p < 0.05); none of the isolates was MDR strains.

Understanding and Exploiting the Effect of Tuberculosis Antimicrobials on Host Mitochondrial Function and Bioenergetics.

Almost 140 years after its discovery, tuberculosis remains the leading infectious cause of death globally. For half a century, patients with drug-sensitive and drug-resistant tuberculosis have undergone long, arduous, and complex treatment processes with several antimicrobials that primarily function through direct bactericidal activity. Long-term utilization of these antimicrobials has been well-characterized and associated with numerous toxic side-effects. With the prevalence of drug-resistant strains on the rise and new therapies for tuberculosis urgently required, a more thorough understanding of these antimicrobials is a necessity. In order to progress from the “one size fits all” treatment approach, understanding how these antimicrobials affect mitochondrial function and bioenergetics may provide further insight into how these drugs affect the overall functions of host immune cells during tuberculosis infection. Such insights may help to inform future studies, instigate discussion, and help toward establishing personalized approaches to using such antimicrobials which could help to pave the way for more tailored treatment regimens. While recent research has highlighted the important role mitochondria and bioenergetics play in infected host cells, only a small number of studies have examined how these antimicrobials affect mitochondrial function and immunometabolic processes within these immune cells. This short review highlights how these antimicrobials affect key elements of mitochondrial function, leading to further discussion on how they affect bioenergetic processes, such as glycolysis and oxidative phosphorylation, and how antimicrobial-induced alterations in these processes can be linked to downstream changes in inflammation, autophagy, and altered bactericidal activity.

Stopping of the Downtrend of Tuberculosis in Iran, a Systematic Review of Associated Risk Factors.

Tuberculosis (TB) is a public health problem in developing countries. In recent decades, the incidence of the disease has been gradually reducing in Iran. However, the reducing incidence of the disease has stopped in the country during recent years. It could be due to an increase in immigration, diabetes, HIV/AIDS, and the prevalence of drug-resistant strains. In order to prevent the spread of TB cases and control this disease, it is essential to identify the predisposing factors, which may be related to bacteria, host and environment. The objective of the present systematic review was to investigate the role of potentially effective factors in the increase in TB cases in the country. The epidemiological studies that had considered the risk factors for the development of TB in populations from different regions of Iran were reviewed systematically from the beginning of 2007 to the end of June 2017 in electronic databases. Upon evaluation of the literature, these 7 major risk factors were identified in twenty-five eligible studies, including poor living conditions, drug abuse, HIV/AIDS, multidrug-resistant tuberculosis (MDR-TB), diabetes, migration, and smoking. In conclusion, the increase in predisposing risk factors for catching TB, especially the migration and Beijing strain, shows that in the absence of accurate monitoring, TB cases will increase in the near future in Iran.

Multi-drug resistant and extended-spectrum \u03b2-lactamases producing bacterial uropathogens among pregnant women in Northwest Ethiopia

IntroductionAbove 80% of urinary tract infections are caused by enteric bacteria, which are known for years by their drug-resistant ability. Though the prevalence of drug-resistant strains is increasing in the world, it is not well known in low-income countries. The aim of this study was to assess the prevalence of Multi-drug resistance, Extended-spectrum β-lactamases production, and associated risk factors among pregnant women in Northwest Ethiopia.MethodsA hospital-based cross-sectional study was conducted among pregnant women from March to May 2017. A total of 384 clean-catch midstream urine sample was collected from study participants. Bacterial identification and drug susceptibility testing were done following standard microbiological techniques; Extended-spectrum β-lactamase production was screened using a disc diffusion test and confirmed by a combination disc test. The data were entered and analyzed by using SPSS version 20, and a p-value of less than 0.05 was considered as statistically significant.ResultThe overall prevalence of urinary tract infection was 15.9% (95% CI 12.8–20.1%). E. coli (49.2%), CoNS (27.9%), and S. aureus (18%) were the main uropathogens. The prevalence of MDR uropathogens was 60.65%. The prevalence of ESBLs production among cases caused by Enterobacteriaceae was 18.2%. The drug resistance rate of Gram-negative isolates was higher for ampicillin (90.9%), cephalothin (84.8%), and augmentin (57.6%). The drug nitrofurantoin showed the highest activity (100%) against Gram-negative isolates. Gram-positive isolates were showed low susceptibility to penicillin (89.3%) and cotrimoxazole (75%); however highest susceptibility rate for gentamicin (100%), amikacin (100%), and nitrofurantoin (98.36%) was recorded. Prior antibiotic therapy (AOR = 5.46, 95% CI 1.38–21.65) was a risk factor for the presence of multi-drug resistant bacteria.Conclusion and recommendationThe multi-drug resistance prevalence was high among uropathogen, thus treatment of urinary tract infection during pregnancy; should be based on the antibacterial susceptibility testing result. The isolation of drug-resistant strains like Extended-spectrum β-lactamases in this study calls for the need of periodic and continuous follow-up of antibiotic usage among pregnant women. Nitrofurantoin, gentamicin, amikacin, and ciprofloxacin/norfloxacin showed higher activity against bacterial uropathogen.

Modelling intra-host competition between malaria parasites strains

An intra-host epidemiological model is formulated for the co-infection of drug-sensitive and drug-resistant malaria parasites to examine the impact of aggressive treatment on the effect of competitive release in an infected host. The analysis of the existence of equilibrium and their stability of the model is conducted, and the results reveal that the intra-host competition and treatment play a key role in the prevalence of drug-resistant strains. The mathematical outcomes qualitatively match the experimental fact, that the rapid elimination of drug-resistant strains could promote the very evolution it is intended to retard.

A barcode of multilocus nuclear DNA identifies genetic relatedness in pre- and post-Artemether/Lumefantrine treated Plasmodium falciparum in Nigeria

BackgroundThe decline in the efficacy of artemisinin-based combination treatment (ACT) in some endemic regions threatens the progress towards global elimination of malaria. Molecular surveillance of drug resistance in malaria-endemic regions is vital to detect the emergence and spread of mutant strains.MethodsWe observed 89 malaria patients for the efficacy of artemether-lumefantrine for the treatment of uncomplicated Plasmodium falciparum infections in Lagos, Nigeria and determined the prevalence of drug resistant strains in the population. Parasite clearance rates were determined by microscopy and the highly sensitive var gene acidic terminal sequence (varATS) polymerase chain reaction for 65 patients with samples on days 0, 1, 3, 7, 14, 21 and 28 after commencement of treatment. The genomic finger print of parasite DNA from pre- and post-treatment samples were determined using 24 nuclear single nucleotide polymorphisms (SNP) barcode for P. falciparum. Drug resistance associated alleles in chloroquine resistance transporter gene (crt-76), multidrug resistance genes (mdr1–86 and mdr1–184), dihydropteroate synthase (dhps-540), dihydrofolate reductase (dhfr-108) and kelch domain (K-13580) were genotyped by high resolution melt analysis of polymerase chain reaction (PCR) fragments.ResultsBy varATS qPCR, 12 (18.5%) of the participants had detectable parasite DNA in their blood three days after treatment, while eight (12.3%) individuals presented with genotypable day 28 parasitaemia. Complexity of infection (CoI) was 1.30 on day 0 and 1.34 on day 28, the mean expected heterozygosity (HE) values across all barcodes were 0.50 ± 0.05 and 0.56 ± 0.05 on days 0 and 28 respectively. Barcode (π) pairwise comparisons showed high genetic relatedness of day 0 and day 28 parasite isolates in three (37.5%) of the eight individuals who presented with re-appearing infections. Crt-76 mutant allele was present in 38 (58.5%) isolates. The mdr1–86 mutant allele was found in 56 (86.2%) isolates. No mutation in the K-13580 was observed.ConclusionsPersistence of DNA-detectable parasitaemia in more than 18% of cases after treatment and indications of genetic relatedness between pre- and post-treatment infections warrants further investigation of a larger population for signs of reduced ACT efficacy in Nigeria.

Potential applications of lactic acid bacteria and bacteriocins in anti-mycobacterial therapy

Tuberculosis (TB) is a communicable disease caused by Mycobacterium tuberculosis (M. tuberculosis). WHO estimated that 10.4 million new (incident) TB cases worldwide in year 2016. The increased prevalence of drug resistant strains and side effects associated with the current anti-tubercular drugs make the treatment options more complicated. Hence, there are necessities to identify new drug candidates to fight against various sub-populations of M. tuberculosis with less or no toxicity/side effects and shorter treatment duration. Bacteriocins produced by lactic acid bacteria (LAB) attract attention of researchers because of its “Generally recognized as safe” status. LAB and its bacteriocins possess an effective antimicrobial activity against various bacteria and fungi. Interestingly bacteriocins such as nisin and lacticin 3147 have shown antimycobacterial activity in vitro. As probiotics, LAB plays a vital role in promoting various health benefits including ability to modulate immune response against various infectious diseases. LAB and its metabolic products activate immune system and thereby limiting the M. tuberculosis pathogenesis. The protein and peptide engineering techniques paved the ways to obtain hybrid bacteriocin derivatives from the known peptide sequence of existing bacteriocin. In this review, we focus on the antimycobacterial property and immunomodulatory role of LAB and its metabolic products. Techniques for large scale synthesis of potential bacteriocin with multifunctional activity and enhanced stability are also discussed.

Drug-Resistant Tuberculosis and Modern Approaches towards its Diagnosis

The review of the literature has shown that tuberculosis as a socially conditioned disease remains a serious problem in Kazakhstan and worldwide in general, currently taking the first position among the infectious diseases causing mortality. One of the main reasons of poor effectiveness of treatment and heavy mortality caused by tuberculosis is prevalence of multiple resistance of tuberculosis mycobacteria to antituberculotics. Modern immunological and molecular genetic tuberculosis diagnostic methods, in comparison with the traditional methods, are more sensitive and specific and allow within a short time detecting the presence of tuberculosis mycobacteria in bio-substrates and their resistance to drugs, which reduces the time of abacillation among the patients, increases effectiveness of the treatment, prevents prevalence of drug-resistant strains of tuberculosis mycobacteria, should allow decreasing the tuberculosis infection rate in general and incidence of Multiple/Extensive Drug-Resistant tuberculosis in particular.

PREVALENCE OF DRUG RESISTANT HIV STRAINS IN HIV-INFECTED PATIENTS OF REPRODUCTIVE AGE

The prevalence of drug resistant HIV strains among HIV-positive reproductive aged persons with ineffective antiretroviral therapy (ART) was assessed. The prevalence of drug resistant strains of HIV was 73.8% in the group of women and 89.29% in the group of men (totally in 80.0% of patients). In the spectrum of drug resistance mutations (DRMs) the most prevalent mutation associated with high-level resistance to nucleoside reverse transcriptase inhibitors was substitution M184V (80.36%); in addition, the high prevalence of K65R (26.79%) was indicated. The most common mutations causing a high-level resistance to nonnucleoside reverse transcriptase inhibitors were G190S/A (57.14%), Y181C (37.50%), K101E (33.93%). The DRMs to protease inhibitors were indicated with significantly less frequent (5.36%).

Prevalence of tuberculosis in prisons: risk factors and molecular epidemiology.

Tuberculosis (TB) in prisons is a challenge for the public health system. Active and passive case screening are important tools for TB case detection. To characterise TB in a southern Brazil prison in terms of epidemiological variables, diagnostic approaches and clinical isolate genotypes. Inmates of a southern Brazilian prison were assessed using active and passive TB case screening. Sputum microscopy, culture, drug susceptibility testing and genotyping were performed. Data were analysed using descriptive statistics and multivariable logistic regression. TB prevalence was 4712 per 100 000 inmates, and was associated with low educational level, time incarcerated, productive cough, previous TB history, smoking and human immunodeficiency virus (HIV) infection. Overall, 27.8% of TB cases were detected by culture only; the prevalence of drug-resistant strains was 7.8%; 58.3% of clinical isolates had an identical genotypic profile. The study showed extensive circulation of Mycobacterium tuberculosis strains in a highly endemic prison. It is recommended that priority be given to the evaluation of prison inmates with longer jail times, those who are HIV-positive, those with symptoms and those with a previous history of tuberculosis. We observed that active case finding induced passive case detection.

Evaluation of sensitivity and specificity of GenoType MTBDRplus line probe assay in rapid diagnosis of tuberculous pleural effusion

BackgroundPakistan has an annual tuberculosis (TB) incidence rate of 233 per 100,000 population and ranks sixth among the high TB burden countries worldwide. Tuberculous pleural effusion (TPE) occurs in up to 25% of TB patients. Owing to the pauci-bacillary nature of the pleural fluid, the diagnosis of TPE is a challenge. ObjectiveThe aim of the present study is to evaluate the performance of MTBDRplus line probe assay in terms of its sensitivity and specificity for the rapid TB diagnosis in TPE and frequency of multidrug-resistant TB (MDR-TB) in TPE through the detection of rifampicin (Rif) and isoniazid (INH) resistance. MethodsThis study is an ongoing prospective cross-sectional study being performed at Aga Khan University Hospital. Patients aged 18years or more with suspected TPE were asked to participate. In addition to a preformed questionnaire which gathered information regarding clinical history and previous laboratory investigations, already tapped pleural effusion samples are tested for pleural fluid composition, TB smear and culture, MTBDRplus line probe assay and ADA analysis. ResultsSo far, 22 patients with a mean age of 49.81±21.75 (range: 18–86; IQR: 29.25, 68.00) years have been enrolled. Almost two-thirds are males (n=15, 68.2%). The majority of the patient population belongs to Karachi (n=18, 81.8%), followed by Quetta (n=2, 9.1%), Faisalabad (n=1, 4.5%) and Sukkur (n=1, 4.5%). Half of the patients had a history of TB contact (n=11, 50.0%), whereas only two (9.1%) had a past history of TB. Fifty percent (n=11) had a right-sided pleural effusion. Effusion size was moderate in 36.4% (n=8), mild in 27.3% (n=6) and massive in 22.5% (n=5). Pleural fluid MTBDRplus positively detected TB in 9.1% (n=2) of cases. INH resistance was detected in one of these two cases. Pleural fluid culture was positive in two cases. Pleural fluid AFB smear was negative in all cases. ConclusionGiven the study is still under way, it is hoped that the results of this study may investigate the relevance, sensitivity and specificity of MTBDRplus line probe assay as a tool in the rapid detection of MTB in TPE and the early detection of MDR-TB through the detection of Rif and INH resistance. It may also provide an estimate of prevalence of drug resistant strains in TPE, which would be useful for guiding empirical anti-tuberculous therapy in TPE.

Using a diabetes drug to treat tuberculosis

Tuberculosis The increasing prevalence of drug-resistant strains of Mycobacterium tuberculosis —the bacterium that causes tuberculosis (TB)—has led to a paradigm shift in the search for new drugs. Rather than targeting the bacterium itself, researchers are trying to augment host defenses. Now,

Rifoligotyping assay: an alternative method for rapid detection of rifampicin resistance in Mycobacterium tuberculosis isolates from Morocco

One of the greatest threats to global tuberculosis (TB) control is the growing prevalence of drug resistant strains. In the past decades, considerable efforts have been made upon the development of new molecular technologies and methodologies for detection of drug resistance in Mycobacterium tuberculosis (MTB). A sensitive, specific reverse line blot assay, called rifoligotyping (RIFO), for the detection of genotypic resistance to rifampicin (RIF), was designed and evaluated. RIFO includes oligonucleotide probes specific for wild-type and mutant sequences, allowing specific and sensitive detection of both genotypes in a single assay. The RIFO was applied on 500 MTB isolates from Morocco. The results of the RIFO showed a good sensitivity (90.9%) and high specificity (100%); the positive and negative predictive values were 100% and 96.1%, respectively. This rapid, simple, economical assay provides a practical alternative for RIF genotyping, especially in low-income countries, to improve TB control and management.