Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis

Ikryannikova, Larisa N.; Afanas’ev, M. V.; Akopian, T. A.; Ilina, Elena; Kuzmin, Alexey V.; Ларионова, Е. Е.; Смирнова, Т. Г.; Черноусова, Л. Н.; Govorun, Vadim M.

doi:10.1016/j.mimet.2007.05.015

Cited by 17 publications

(10 citation statements)

References 34 publications

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“…Because the majority of resistance mechanisms in N. gonorrhoeae are linked to mutations in the chromosomal DNA, adequate methods for SNP scanning should be created. MALDI-TOF mass spectrometry-based minisequencing or a sequencing procedure for the detection of SNPs was applied successfully for hepatitis C virus genotyping (13), detection of genetic markers of antibacterial resistance in Mycobacterium tuberculosis (12), and discrimination of mef genes in Streptococcus pneumoniae (unpublished data). This approach has been applied to N. gonorrhoeae strains for the first time.…”

Section: Discussionmentioning

confidence: 99%

Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains

Ilina

Vereshchagin

Borovskaya

et al. 2008

Antimicrob Agents Chemother

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The main goal of this work is to clarify the predictive value of known genetic markers of Neisseria gonorrhoeae resistance to penicillin, tetracycline, and fluoroquinolones. The correlation between the presence of certain genetic markers and susceptibility of N. gonorrhoeae isolates to penicillin, tetracycline, and fluoroquinolones has been analyzed by means of statistical methods. Susceptibility testing with penicillin, tetracycline, and fluoroquinolones was performed by the agar dilution method. N. gonorrhoeae genomic DNA was isolated. The presence of bla TEM-1 and tet(M) genes was analyzed by PCR. A novel method of polymorphism discovery based on a minisequencing reaction followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry was applied for the analysis of chromosomal N. gonorrhoeae genes involved in antimicrobial resistance development. Clinical N. gonorrhoeae isolates (n ‫؍‬ 464) were collected. Susceptibility levels to penicillin, tetracycline, and fluoroquinolones were found to be 25.9%, 35.9%, and 54.1%, respectively. Among the 19 N. gonorrhoeae isolates with penicillin MICs of >4 g/ml, the bla TEM-1 gene was detected in 12. The Tet(M) determinant was found in 4 of 12 N. gonorrhoeae isolates with tetracycline MICs of >16 g/ml. The chromosomal genetic markers of penicillin and tetracycline resistance were detected especially in isolates with penicillin MICs of 0.25 to 2.0 g/ml and tetracycline MICs of 0.5 to 4 g/ml. Mutations in GyrA and ParC were found in 208 of 211 quinoloneresistant N. gonorrhoeae isolates. This work is the first representative molecular research of the N. gonorrhoeae population in Russia. Information about the prevalence of antibiotic resistance mechanisms and the positive predictive value of certain genetic determinants is given. The positive predictive values of the analyzed genetic markers were found to be different for fluoroquinolones (90.3%), penicillin (91.1%), and tetracycline (81.9%).

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Section: Discussionmentioning

confidence: 99%

Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains

Ilina

Vereshchagin

Borovskaya

et al. 2008

Antimicrob Agents Chemother

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“…Перспективное направление научных исследований связано с таргетным секвенированием генов для поиска основ лекарственной резистентности МБТ, особенно к резервным противотуберкулезным препаратам, а также различных видов нетуберкулезных микобактерий [23][24][25][26][27].…”

Section: микробиология и молекулярная генетика микобактериальных инфеunclassified

Tuberculosis in the Russian Federation: Situation, Challenges and Perspectives

Ergeshov¹

2018

Annals RAMS

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Tuberculosis (TB) is a significant problem of public health both in Russia and abroad. About one third of the world’s population is infected with Mycobacterium tuberculosis. Every year more than 10 million new TB cases are registered in the world, and about 1.7 million people die from TB. In the Russian Federation, due to the measures taken by the Government and health authorities the epidemic TB situation has been noticeably improved since the sharp deterioration in the 90s of the last century. At the same time, the spread of drug-resistant TB and its low treatment effectiveness, the spread of combined HIV and TB infection reduces the effectiveness of anti-tuberculosis interventions. The research conducted by CTRI, the WHO Collaborating Center for TB in the Russian Federation, is aimed at solving such urgent problems as studying latent infection mechanisms, developing new test systems for accelerated diagnostics of drug resistance, clinical approbation and introduction of short effective regimens of chemotherapy, developing new antituberculosis agents. The achieved successes in the study of tuberculosis in the 21st century creates preconditions for eliminating its epidemic both in Russia and the world, though suggesting imminent breakthrough for tuberculosis is a hasty conclusion.

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“…Resistance to rifampin and isoniazid in M. tuberculosis (67,68) was also analyzed by MALDI-TOF MS sequencing.…”

Section: Minisequencingmentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) Mass Spectrometry for Detection of Antibiotic Resistance Mechanisms: from Research to Routine Diagnosis

2013

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SUMMARYMatrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied as an identification procedure in clinical microbiology and has been widely used in routine laboratory practice because of its economical and diagnostic benefits. The range of applications of MALDI-TOF MS has been growing constantly, from rapid species identification to labor-intensive proteomic studies of bacterial physiology. The purpose of this review is to summarize the contribution of the studies already performed with MALDI-TOF MS concerning antibiotic resistance and to analyze future perspectives in this field. We believe that current research should continue in four main directions, including the detection of antibiotic modifications by degrading enzymes, the detection of resistance mechanism determinants through proteomic studies of multiresistant bacteria, and the analysis of modifications of target sites, such as ribosomal methylation. The quantification of antibiotics is suggested as a new approach to study influx and efflux in bacterial cells. The results of the presented studies demonstrate that MALDI-TOF MS is a relevant tool for the detection of antibiotic resistance and opens new avenues for both clinical and experimental microbiology.

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Mass-spectrometry based minisequencing method for the rapid detection of drug resistance in Mycobacterium tuberculosis

Cited by 17 publications

References 34 publications

Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains

Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains

Tuberculosis in the Russian Federation: Situation, Challenges and Perspectives

Matrix-Assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) Mass Spectrometry for Detection of Antibiotic Resistance Mechanisms: from Research to Routine Diagnosis

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