Spoliogotyping - A key to Genetic insights

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 Spoligotyping

  • Spoligotyping is a molecular biological method used to describe the genetic diversity of Mycobacterium tuberculosis complex (MTBC) species in humans and animals.
  • It involves amplifying a specific DNA region called the direct repeat (DR) region using oligonucleotide primers and then hybridizing the PCR product to a set of immobilized oligonucleotides corresponding to unique spacer DNA sequences within the DR locus.
  • The results are interpreted by converting black squares to 1 and white squares to 0, which are then transferred to databases for further analysis.
  • Spoligotyping helps identify spoligotype patterns, lineages, and international types of MTBC isolates, providing valuable information for understanding TB epidemiology and transmission dynamic.



            Fig. 1: Genetic characterization of Mycobacterium tuberculosis strains from different regions

The figure presents a dendrogram showing the genetic relatedness of various Mycobacterium tuberculosis strains based on spoligotyping and RFLP-IS6110 patterns. Each strain is identified by a specific identifier (e.g., VEZ 399, VEZ 479) and its corresponding geographic state of isolation. The presence (1) or absence (0) of mutations in the katG and rpoB genes, which are associated with resistance to the antibiotics isoniazid and rifampicin, respectively, are indicated. The Shared International Type (SIT) number classifies the strains according to a global database. The clustering in the dendrogram illustrates the genetic similarity among the strains, highlighting the genetic diversity and the spread of antibiotic resistance.


Case Study

  • Spoligotyping was used in a cross-sectional study in the Oromia region of Ethiopia to investigate Mycobacterium tuberculosis complex (MTBC) species circulating in the population .
  • The study recruited 450 smear positive pulmonary TB cases and isolated mycobacteria from sputum samples for molecular identification using spoligotyping .
  • Results revealed 315 isolates grouped under 181 different spoligotype patterns, with the most dominant pattern being SIT149 comprising 23 isolates .
  • The majority of isolates belonged to Euro-American (EA), East-African-Indian (EAI), and Indo-Oceanic (IO) lineages, with clustering associated with the EAI lineage .
  • The study highlighted the predominance of certain spoligotypes and lineages in the region, emphasizing the need for further genetic typing methods to enhance understanding of TB transmission dynamics .

Reference - doi:10.3389/fpubh.2022.808626


Behind "The Biotechnology Journal"
Mansi Popat & Japan Raval


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