Insertion- and Deletion-Associated Genetic Diversity of Mycobacterium tuberculosis Phospholipase C-Encoding Genes among 106 Clinical Isolates from Turkey

The Mycobacterium tuberculosis genome contains four highly related genes which present significant similarity to Pseudomonas aeruginosa genes encoding phospholipase C enzymes. Three of these genes, plcA, plcB and plcC, are organized in tandem (locus plcABC). The fourth gene, plcD, is located in a different region. This study investigates variations in plcABC and plcD genes in clinical isolates of M. tuberculosis, Mycobacterium africanum and ‘Mycobacterium canettii’. Genetic polymorphisms were examined by PCR, Southern blot hybridization, sequence analysis and RT-PCR. Seven M. tuberculosis isolates contain insertions of IS6110 elements within plcA, plcC or plcD. In 19 of 25 M. tuberculosis isolates examined, genomic deletions were identified, resulting in loss of parts of genes or complete genes from the plcABC and/or plcD loci. Partial plcD deletion was observed in one M. africanum isolate. In each case, deletions were associated with the presence of a copy of the IS6110 element and in all occurrences IS6110 was transposed in the same orientation. A mechanism of deletion resulting from homologous recombination of two copies of IS6110 was recognized in a group of genetically related M. tuberculosis isolates. Five M. tuberculosis isolates presented major polymorphisms in the plcABC and plcD regions, along with loss of expression competence that affected all four plc genes. Phospholipase C is a well-known bacterial virulence factor. The precise role of phospholipase C in the pathogenicity of M. tuberculosis is unknown, but considering the potential importance that the plc genes may have in the virulence of the tubercle bacillus, the study of isolates cultured from patients with active tuberculosis bearing genetic variations affecting these genes may provide insights into the significance of phospholipase C enzymes for tuberculosis pathogenicity.

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Corrigendum to ‘Genetic diversity and major spoligotype families of drug-resistant Mycobacterium tuberculosis clinical isolates from different regions of Turkey’ [Inf. Gen. Evol. 7 (2007) 513–519]

Infection Genetics and Evolution ◽

10.1016/j.meegid.2007.09.004 ◽

2008 ◽

Vol 8 (2) ◽

pp. 227

Author(s):

Riza Durmaz ◽

Thierry Zozio ◽

Selami Gunal ◽

Akgun Yaman ◽

Cengiz Cavusoglu ◽

...

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Clinical Isolates ◽

Drug Resistant

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Characterization of the Genetic Diversity of Extensively-Drug Resistant Mycobacterium tuberculosis Clinical Isolates from Pulmonary Tuberculosis Patients in Peru

PLoS ONE ◽

10.1371/journal.pone.0112789 ◽

2014 ◽

Vol 9 (12) ◽

pp. e112789 ◽

Cited By ~ 13

Author(s):

Omar Cáceres ◽

Nalin Rastogi ◽

Carlos Bartra ◽

David Couvin ◽

Marco Galarza ◽

...

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Pulmonary Tuberculosis ◽

Clinical Isolates ◽

Drug Resistant ◽

Tuberculosis Patients ◽

Extensively Drug Resistant

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Assessment of Mycobacterial Interspersed Repetitive Unit-QUB Markers To Further Discriminate the Beijing Genotype in a Population-Based Study of the Genetic Diversity of Mycobacterium tuberculosis Clinical Isolates from Okinawa, Ryukyu Islands, Japan

Journal of Clinical Microbiology ◽

10.1128/jcm.00348-07 ◽

2007 ◽

Vol 45 (11) ◽

pp. 3606-3615 ◽

Cited By ~ 18

Author(s):

J. Millet ◽

C. Miyagi-Shiohira ◽

N. Yamane ◽

C. Sola ◽

N. Rastogi

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Population Based ◽

Clinical Isolates ◽

Beijing Genotype ◽

Ryukyu Islands ◽

Population Based Study

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Molecular Investigation of hemolytic phospholipase C Encoding Genes in Clinical Isolates of Pseudomonas aeruginosa

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9ispl1.1387 ◽

2018 ◽

Vol 9 (SPL1) ◽

Author(s):

Rasha Hadi Saleh ◽

Habeeb S Naher ◽

Mohammed AK Al-saadi

Keyword(s):

Phospholipase C ◽

Virulence Genes ◽

Clinical Isolates ◽

Clinical Samples ◽

Burn Wound ◽

Molecular Investigation ◽

Genes Encoding ◽

Stool Samples ◽

Polymerase Chain ◽

Encoding Genes

This study is aimed to isolate P.aeruginosa from different clinical cases and to detect the prevalence of virulence genes encoding hemolytic phospholipase C(plcH)in these clinical isolates. In this study a total of 422 clinical samples including burn,wound,ear,urine,abscess and stool were aseptically taken from out- and inpatients who admitted into two hospitals in Hilla City (Teaching Al-Hilla Hospital and Babylon Hospital for Maternity and children during a period of three months. All samples were subjected to bacterial cultivation for the isolation of P.aeruginosa. The isolated P.aeruginosa was diagnosed depended on morphological,biochemical and molecular standard characteristics. Hemolytic phospholipase Cencoding genes(plcH) were detected by PCR and the amplification products were separated in 1% agarose gels containing ethidium bromide. Out of 422 samples,P.aeruginosa was isolated from 54 samples (12.8%). The distribution of these isolates were: 22 (55%) from burn samples,2; (50%) from diabitics foot samples,8 (14.8%) from wound samples, 8 (32%) from ear samples,3 (11%) from abscess samples, 7 (4%) from stool samples,4 (4%) from urine samples and 0 sputum samples. The genotypic properties of hemolytic phospholipase C (plcH )toxins was detected by polymerase chain reaction (PCR). The results of this study revealed that(plcH )gene found in 13/20 (65%)of isolates.

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Genetic diversity of Mycobacterium tuberculosis clinical isolates in Blantyre, Malawi

Heliyon ◽

10.1016/j.heliyon.2019.e02638 ◽

2019 ◽

Vol 5 (10) ◽

pp. e02638 ◽

Cited By ~ 2

Author(s):

Victor Ndhlovu ◽

Anmol Kiran ◽

Derek Sloan ◽

Wilson Mandala ◽

Konstantina Kontogianni ◽

...

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Clinical Isolates

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In Vitro Activity of β-Lactams in Combination with β-Lactamase Inhibitors against Mycobacterium tuberculosis Clinical Isolates

BioMed Research International ◽

10.1155/2018/3579832 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Fu Li ◽

Li Wan ◽

Tongyang Xiao ◽

Haican Liu ◽

Yi Jiang ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Targets ◽

Synergetic Effect ◽

Drug Susceptibility ◽

Clinical Isolates ◽

Nucleotide Sequencing ◽

Drug Susceptibility Test ◽

Single Target ◽

Encoding Genes

Objectives. Evaluating the activity of nineteen β-lactams in combination with different β-lactamase inhibitors to determine the most potent combination against Mycobacterium tuberculosis (MTB) in vitro. Methods. Drug activity was examined by drug susceptibility test with 122 clinical isolates from China. Mutations of blaC and drug targets ldtMt1, ldtMt2, dacB2, and crfA were analyzed by nucleotide sequencing. Results. Tebipenem (TBM) in combination with clavulanate (CLA) exhibited the highest anti-TB activity. The MIC of β-lactam antibiotics was reduced most evidently in the presence of CLA, compared to avibactam (AVI) and sulbactam (SUB). Eight polymorphism sites were identified in blaC, which were not associated with β-lactams resistance. Interestingly, one strain carrying G514A mutation in blaC was highly susceptible to β-lactams regardless of the presence of inhibitors. The transpeptidase encoding genes, ldtMt1, ldtMt2, and dacB2, harboured three mutations, two mutations, and one mutation, respectively, but no correlation was found between these mutations and drug resistance. Conclusion. The activity of β-lactams against MTB and different synergetic effect of β-lactamase inhibitors were indicated. TBM/CLA exhibited the most activity and has a great prospect in developing novel anti-TB regimen; however, further clinical research is warranted. Moreover, the resistance to the β-lactam antibiotics might not be conferred by single target mutation in MTB and requires further studies.

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Enterobacterial Repetitive Intergenic Consensus Sequences as Molecular Targets for Typing of Mycobacterium tuberculosis Strains

Journal of Clinical Microbiology ◽

10.1128/jcm.36.1.128-132.1998 ◽

1998 ◽

Vol 36 (1) ◽

pp. 128-132 ◽

Cited By ~ 20

Author(s):

Leonardo A. Sechi ◽

Stefania Zanetti ◽

Ilaria Dupré ◽

Giovanni Delogu ◽

Giovanni Fadda

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Molecular Targets ◽

Clinical Isolates ◽

Gram Negative Bacteria ◽

Enterobacterial Repetitive Intergenic Consensus ◽

Gram Negative ◽

Consensus Sequences ◽

First Time

The presence of enterobacterial repetitive intergenic consensus (ERIC) sequences was demonstrated for the first time in the genome ofMycobacterium tuberculosis; these sequences have been found in transcribed regions of the chromosomes of gram-negative bacteria. In this study genetic diversity among clinical isolates of M. tuberculosis was determined by PCR with ERIC primers (ERIC-PCR). The study isolates comprised 71 clinical isolates collected from Sardinia, Italy. ERIC-PCR was able to identify 59 distinct profiles. The results obtained were compared with IS6110 and PCR-GTG fingerprinting. We found that the level of differentiation obtained by ERIC-PCR is greater than that obtained by IS6110fingerprinting and comparable to that obtained by PCR-GTG. This method of fingerprinting is rapid and sensitive and can be applied to the study of the epidemiology of M. tuberculosis infections, especially when IS6110 fingerprinting is not of any help.

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