26Beijing strains of Mycobacterium tuberculosis (lineage2) have been associated 27 with drug-resistance and transmission of tuberculosis worldwide. Most of the Beijing 28 strains identified in the Colombian pacific coast exhibited a multidrug resistant (MDR) 29 phenotype, in contrast with the phenotype observed in Beijing isolates from other South-30 American countries. We wanted to evaluate the clonality and genetic background of the 31 Beijing strains isolated in Colombia that belong mostly to the spoligo-international type 32 (SIT) 190. Out of 37 Beijing stains characterized in an 8-years period, we identified five 33 Beijing clones; 36 that belong to the SIT190 type and only one to SIT1. Two loci in 34 VNTR typing: MIRU 39 and QUB11b exhibited the highest level of variation among 35 these strains. Out of the 37 Beijing strains, only one was drug susceptible, 28 36 represented MDR-TB, four extensively drug resistant TB (XDR-TB) and four pre-XDR-37 TB. The mutations rpoB S531L and katG S315T1 were the most common among the 38 MDR strains as reported elsewhere. Whole genome sequencing analysis allowed us to 39 classify them as modern lineage Beijing strains, sharing up to 76 out of the 275 SNPs 40 described in Beijing strains, as identified worldwide by Schürch et al; including 54 non-41 synonymous SNPs and 23 silent mutations. We were also able to confirm the presence 42 of 8 specific SNPs that were so far only found in the Beijing strains from Colombia. The 43 presence of modern Beijing strains, most of them representing MDR-TB, suggests a 44 different origin of this M. tuberculosis lineage compared to other Beijing strains found in 45 neighboring countries, such as Peru. The specific 8-SNP signature confirmed the 46 identity of these Colombian Beijing MDR strains. This work may serve as a genetic 3 47 baseline to study the evolution and spread of M. tuberculosis Beijing strains in 48 Colombia, which play an important role in the control of MDR-TB.
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