Utility of mycobacterial interspersed repetitive unit typing for differentiating Mycobacterium tuberculosis isolates in Wuhan, China

Mycobacterial interspersed repetitive unit (MIRU) typing has been found to allow rapid, reliable, high-throughput genotyping of Mycobacterium tuberculosis, and may represent a feasible approach to study M. tuberculosis molecular epidemiology. To evaluate the use of MIRU typing in discriminating M. tuberculosis strains, isolates from 105 patients in Wuhan City, China, were genotyped by this method as compared to spoligotyping. MIRU typing identified 55 types that defined 21 clusters and 34 unique isolates. The discriminatory power was high [Hunter–Gaston discriminatory index (HGDI), 0.97]. Spoligotyping showed that 86 (81.9 %) of 105 isolates belonged to the Beijing family genotype. For Beijing family and non-Beijing strains, the discriminatory power of MIRU was high (HGDI, 0.95 and 0.98, respectively). Among the alleles of the MIRU loci for the Beijing family, only locus 26 was highly discriminative, but for non-Beijing strains, loci 10, 16 and 26 were highly discriminative. MIRU typing is a simple and fast method which may be used for preliminary screening of M. tuberculosis isolates in China.

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Genotypes of Mycobacterium tuberculosis isolates circulating in Shaanxi Province, China

PLoS ONE ◽

10.1371/journal.pone.0242971 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0242971

Author(s):

Yan Li ◽

Yu Pang ◽

Tianhua Zhang ◽

Xiaoping Xian ◽

Jian Yang ◽

...

Keyword(s):

Genetic Diversity ◽

Drug Resistance ◽

Mycobacterium Tuberculosis ◽

Multidrug Resistant ◽

Shaanxi Province ◽

Drug Resistant ◽

Beijing Family ◽

Tuberculosis Patients ◽

Smear Positive Pulmonary Tuberculosis ◽

Discriminatory Index

Objectives The prevalence of drug-resistant TB in Shaanxi Province is higher than other areas. This study was aimed to investigate the genetic diversity and epidemiology of Mycobacterium tuberculosis clinical strains in Shaanxi Province, China. Methods From January to December 2016, a total of 298 Mycobacterium tuberculosis clinical isolates from smear-positive pulmonary tuberculosis patients were genotyped by Mcspoligotyping and 15-locus VNTR. Results We found that the Beijing family strains was the most prominent family(81.54%, 243/298). Other family strains included T family(9.06%, 27/298), U family(0.67%, 2/298), LAM9 family(0.34%, 1/298) and Manu family(0.34%, 1/298). The rates of multidrug-resistant (MDR) M.Tuberculosis, age, type of case and education between Beijing and non-Beijing family strains were not statistically different, while the distribution in the three different regions among these was statistically significant. VNTR results showed that strains were classified into 280 genotypes, and 33 (11.07%) strains could be grouped into 14 clusters. 11 of the 15-VNTR loci were highly or moderately discriminative according to the Hunter-Gaston discriminatory index. Conclusions We concluded that the Beijing family genotype was the most prevalent genotype and 15-locus VNTR typing might be suitable for genotyping of M. tuberculosis in Shaanxi Province. There was less association between Beijing family genotypes and drug resistance in our study area.

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Application of FLiP Method for Differentiation of Mycobacterium tuberculosis Strains in Comparison to Commonly Used Methods, Spoligotyping and MIRU-VNTR Typing

Polish Journal of Microbiology ◽

10.33073/pjm-2013-009 ◽

2013 ◽

Vol 62 (1) ◽

pp. 73-76 ◽

Cited By ~ 4

Author(s):

ANNA ŻACZEK ◽

ANNA BRZOSTEK ◽

ALINA GÓRNA ◽

ANNA SAJDUDA ◽

ARKADIUSZ WOJTASIK ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Gold Standard ◽

Epidemiological Studies ◽

The Other ◽

Discriminatory Power ◽

Valuable Alternative ◽

Typing Methods ◽

High Level ◽

Discriminatory Index ◽

Vntr Analysis

The current "gold standard" in molecular epidemiological studies of Mycobacterium tuberculosis is IS6110 RFLP based on IS6110 polymorphism. However PCR-based methods are becoming increasingly important. Recently, fast ligation-mediated PCR (FLiP), based on IS6110 polymorphism was proposed. In this study, the discriminatory power of FLIP, spoligotyping and MIRU-VNTR typing, in differentiation of M. tuberculosis isolates was compared. The discriminatory index (HGI) of spoligotyping, MIRU-VNTR analysis, and FLiP was 0.653, 0.837, and 0.917, respectively. This indicates that FLiP allows a high level of differentiation among M. tuberculosis strains and it might be a valuable alternative to the other typing methods.

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A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

BMC Microbiology ◽

10.1186/s12866-021-02212-3 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Sadaf Kalsum ◽

Blanka Andersson ◽

Jyotirmoy Das ◽

Thomas Schön ◽

Maria Lerm

Keyword(s):

Mycobacterium Tuberculosis ◽

High Throughput ◽

High Throughput Screening ◽

Live Cell Imaging ◽

Cell Imaging ◽

Imaging System ◽

Aggregate Size ◽

Live Cell ◽

Screening Assay ◽

High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

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