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Volume 71, Issue 10

Fingerprinting of isolates by MIRU-VNTR genotyping and detection of isoniazid resistance by real-time PCR No Access

Abstract

Tuberculosis (TB) is a great public health problem in developing countries such as Egypt. Genotyping of isolates has a prominent role in the field of TB prevention.

This study aimed to evaluate real-time PCR using Minor Groove Binder (MGB) probes and to identify circulating lineages/sub-lineages of and their transmission patterns.

We hypothesize that MIRU-VNTR technique is efficient in identifying circulating lineages in Egypt.

Fifty sputum specimens positive for acid-fast bacilli were included. Isoniazid (INH) resistance was detected using the 1 % proportion method. Real-time PCR using MGB-probes was used for simultaneous detection of TB infection and INH resistance. Partial sequencing of the gene was used to confirm INH resistance results. A standard 15 Mycobacterial Interspersed Repetitive Unit Variable Number Tandem Repeat (15-MIRU-VNTR) approach was used for genotyping through the MIRU-VNTR online platform.

Only seven specimens showed phenotypic resistance to INH. was detected in all samples, while a mutation in the gene codon 315 was detected only in five samples, which were also phenotypically INH-resistant. Sequencing of the gene showed codon 315 mutation genotypically and phenotypically in the five INH-resistant isolates. Molecular genotyping of isolates revealed that the majority of isolates (26/50, 52 %) belonged to the S family of lineage_4. A low clustering rate (2 %) was observed among our isolates. According to the Hunter-Gaston Discriminatory Index (HGDI), 11 MIRU-VNTR loci were highly or moderately discriminative, while four loci were less polymorphic.

MIRU-VNTR genotyping revealed a low clustering rate with a low recent transmission rate of strains in Alexandria, Egypt.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): genotyping , INH resistance , katG gene , MIRU-VNTR and Mycobacterium tuberculosis
© 2022 The Authors
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2022-10-28
2024-05-01
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Fingerprinting of Mycobacterium tuberculosis isolates by MIRU-VNTR genotyping and detection of isoniazid resistance by real-time PCR
J. Med. Microbiol. 71, 001603 (2022); https://doi.org/10.1099/jmm.0.001603
/content/journal/jmm/10.1099/jmm.0.001603
/content/journal/jmm/10.1099/jmm.0.001603
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