Diagnosis of Mycobacterium tuberculosis using molecular biology technology

Labrador, Jorge; Garberi, Federico; Garberi, Juan; Peneipil, Julián; Garberi, Miguel; Scigliano, Luis; Troncoso, Alcides

doi:10.1016/s2221-1691(11)60002-6

Cited by 24 publications

(8 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 11 These molecular-based assays can be efficiently applied for the rapid identification of genetic mutations and single-nucleotide polymorphisms (SNPs) associated with antibiotic resistance in MTB isolates. 12 Molecular sequencing of genes conferring resistance has been recommended as a reference standard for DST in recent years; however, despite the reliability of the tool, it is costly and may not be easily available in all microbiological laboratories. 5 , 13 High-resolution melting (HRM) curve is a simple, rapid, and accurate technique for scanning of mutations, which relies on the sequence variation that leads to changes in the shape of the melting curve.…”

Section: Introductionmentioning

confidence: 99%

Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of <em>Mycobacterium tuberculosis</em> by high-resolution melting analysis

Sirous¹,

Khosravi²,

Tabandeh³

et al. 2018

IDR

View full text Add to dashboard Cite

BackgroundThe emergence of drug resistance among Mycobacterium tuberculosis (MTB) strains is a serious health concern worldwide. The development of rapid molecular diagnostic methods in recent years has a significant impact on the early detection of resistance to major anti-TB drugs in MTB isolates, which helps in employing appropriate treatment regimen and prevents the spread of drug-resistant strains. This study was designed to evaluate the efficacy of real-time PCR and high-resolution melting (HRM) curve analysis for the determination of resistance to rifampin (RIF), isoniazid (INH), and ofloxacin (OFX) in MTB isolates and to investigate their resistance-related mutations.MethodsHRM analysis was performed to screen 52 (32 drug-resistant and 20 fully susceptible) MTB clinical isolates for mutations in rpoB, katG, mab-inhA, and gyrA genes. The HRM results were then confirmed by DNA sequencing.ResultsIn total, 32 phenotypically resistant isolates, comprising 18 RIF-, 16 INH-, and five OFX- resistant strains, were investigated. HRM analysis successfully identified 15 out of 18 mutations in rpoB, 14 out of 16 mutations in katG and mab-inhA, and four out of five mutations in gyrA conferring resistance to RIF, INH, and OFX, respectively. The obtained sensitivity and specificity, respectively, for HRM in comparison with phenotypic susceptibility testing were found to be 83.3% and 100% for RIF, 87.5% and 100% for INH, and 80% and 100% for OFX. In five resistant strains (12.8%), no mutation was detected by using HRM and DNA sequencing.ConclusionHRM assay is a rapid, accurate, and cost-effective method possessing high sensitivity and specificity for the determination of antibiotic resistance among MTB clinical isolates and screening of their associated mutations. This method can generate results in a shorter period of time than taken by the phenotypic susceptibility testing and also allows for timely treatment and prevention of the emergence of possible MDR strains.

show abstract

Section: Introductionmentioning

confidence: 99%

Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of <em>Mycobacterium tuberculosis</em> by high-resolution melting analysis

Sirous¹,

Khosravi²,

Tabandeh³

et al. 2018

IDR

View full text Add to dashboard Cite

show abstract

“…Because, people need speedy response with high accuracy level diagnosis since the positive impact of test results for infection control decisions towards patient isolation and other therapeutic managements [93]. An accurate TB molecular diagnostic method can be obtained in fewer hours, with an increased sensitivity & specificity [94,95]. Consequently, results will be obtained very shortly than classical methods that favor a timely treatment of the disease that possibly decrease multi-drug resistance development.…”

Section: Increases Precisionsmentioning

confidence: 99%

Molecular Characterization of Mycobacterium bovis and its Significance: Role for Control of Zoonotic Tuberculosis in Africa

Getahun¹

2017

J Med Diagn Meth

View full text Add to dashboard Cite

Mycobacterium bovis is the main causal agent of bovine tuberculosis (BTB) that causes zoonotic tuberculosis (TB) in humans, even though M. caprae; contribute to a lesser extent, which is mostly acquired from domestic animals and their products, in which cattle are the major reservoir. This review paper discussed the importance of zoonotic tuberculosis in Africa and highlighted the uses of molecular methods in diagnosis, control and prevention of zoonotic tuberculosis. Despite the fact, there are various diagnostic methods available nowadays; the molecular techniques are proving more rapid and plausible sensitivity and specificity results that increase the diagnosis precision. In addition, molecular diagnostic methods used to indicate the epidemiological status of different strain distribution with regard to cluster distribution and reactivation of dormant or treated cases of tuberculosis indicating the drug resistance situation of the strain. Hence, applying molecular techniques for diagnosis of zoonotic tuberculosis in developing countries will help in decreasing the incidence, prevalence, mortality, morbidity and loss of economic due to carcass condemnation, milk drop and livestock trade banning in the continent. Advanced molecular methods truly play a key role in prevention and control program of tuberculosis in general and bovine tuberculosis in specific, which leads to the cooperation of various disciplines. Introducing cost effective and easy molecular diagnostic methods is recommended, implementing molecular diagnostic methods in research areas and intensive training of personnel in the use of equipment.

show abstract

“… 3 In addition, the increased incidence of TB is related to the human immune-deficiency virus (HIV) epidemic, which impairs the human immune system. 4 To control the rapid growth of TB and ensure a superior therapeutic effect, combination drugs and fixed dose combinations (FDCs) have become increasingly important. Among these FDCs, pyrazinamide (PZA) and isoniazid (INH) are two typical active pharmaceutical ingredients (APIs) along with rifampicin and ethambutol, which are the most effective drugs in treating TB.…”

Section: Introductionmentioning

confidence: 99%

Vibrational Spectroscopic Investigation into Novel Ternary Eutectic Formed between Pyrazinamide, Fumaric Acid, and Isoniazid

et al. 2020

View full text Add to dashboard Cite

To improve the efficacy of anti-tuberculosis (anti-TB) therapy, drug–drug co-crystallization stands for an alternative approach to settle the tuberculosis problem. Directly co-crystallizing two typical parent anti-TB drugs (pyrazinamide, PZA and isoniazid, INH) into a single binary co-crystal could not be obtained successfully. Multicomponent eutectic are highly effective and useful for enhancing the dissolution rate, bioavailability, and physical stability of the poorly water-soluble active pharmaceutical ingredient (API) drugs, when the attempts of forming a binary co-crystal have failed. Therefore, the ternary eutectic composition conception was proposed in this study, in which fumaric acid (FA) was chosen as the molecule to connect two first-line anti-tubercular drugs. First of all, three starting materials (including PZA, INH, and FA) were grinded at a 1:1:1 molar ratio, the eutectic composition was investigated through vibrational spectroscopic techniques, including terahertz time-domain spectroscopy (THz-TDS) and Raman spectroscopy. Additionally, the density functional theory (DFT) was utilized to simulate the optimized structures and vibrational modes of two possible theoretical eutectic composition forms. The THz absorption spectrum of the theoretical form I shows much more consistency with the experimental results than that of form II. Raman spectra also help to characterize the differences in vibrational modes between the eutectic composition and the starting parent compounds. The results provide us with both structural information and intermolecular hydrogen bonding interactions within specific multicomponent eutectic composition formulations based on Raman and terahertz vibrational spectroscopic techniques in combination with theoretical calculations.

show abstract

Diagnosis of Mycobacterium tuberculosis using molecular biology technology

Cited by 24 publications

References 22 publications

Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of <em>Mycobacterium tuberculosis</em> by high-resolution melting analysis

Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of <em>Mycobacterium tuberculosis</em> by high-resolution melting analysis

Molecular Characterization of Mycobacterium bovis and its Significance: Role for Control of Zoonotic Tuberculosis in Africa

Vibrational Spectroscopic Investigation into Novel Ternary Eutectic Formed between Pyrazinamide, Fumaric Acid, and Isoniazid

Contact Info

Product

Resources

About