They were studied to settle on their LAM family membership. All of them except two (SIT 284 and other with no SIT assigned) presented the LXH254 nmr LAM specific SNP in Ag85C103(GAG→GAA). In addition, we found that two among the isolates tested, or five considering all the
LAM strains, contained the RDRio deletion, which is a feature of a subgroup of the LAM family strains. SCG-6a included a total of 14 isolates, which belonged to T1 (SIT 53, 154, 167, 358, 1122), T2 (SIT 52), T5 (SIT 44), T5_MAD2 (SIT 58), U (SIT 602 and 773) and 4 isolates with not SIT assigned. None of them had either the SNP in Ag85C103 or the SNP in mgtC 182 . This SCG-6a included the isolate of the most representative cluster in 2010, ARA7 (SIT 773, U family), which gathered 133 clinical cases since 2004 [22]. Finally, two unrelated and different isolates presented the same new pattern named SCG-6c, which only differs from SCG-6a in one SNP (Table 2). The first isolate (SIT 90, U) was related with the outbreak ARA21 (20 cases collected since 2004) and the second isolate (SIT 120, T1 family) had not been previously reported
in our Region. Neither contained the SNP in Ag85C103 nor the SNP in mgtC 182 feature for LAM or Haarlem families respectively. Discussion The Euro-American lineage was found to Ralimetinib ic50 be the predominant lineage of the M. tuberculosis complex in Europe [19]. The MDR TB studies carried out in Spain showed the Euro-American as the more prevalent lineage [23], and that a few LAM and Haarlem strains, which belong to this lineage, played a major role in the spread of MDR strains [24]. According to this, the 90% of the tuberculosis strains analysed in this work belong to this lineage. Non-specific serine/threonine protein kinase Our work allowed to classify a collection of MTC strains previously analysed by Spoligotyping and RFLP in Aragon in lineages as well as in SCGs by the detection of the 9 SNPs that define the 7 SCGs [15, 16] together with PCR identification of katG463, Ag85C103 and mgtC182 polymorphisms. All these single polymorphisms as a whole have proved to be an effective complement for both Spoligotyping and RFLP techniques that enhance
their sensibility, especially in those families identified at the beginning as T, U and orphan. A notorious circumstance to remark in our population was that the two largest clusters of M. tuberculosis strains, named ARA21 and ARA7, belonged to T and unclassified groups of families. Besides, ARA7 had caused an outbreak since 2004, what resulted in around the 20% of cases of tuberculosis [22]. This fact allows the classification of these strains into more resolved families. In addition, the 9 SNPs detection by using a pyrosequencing assay leads to obtain quick and PLX3397 datasheet reliable results at an affordable cost [20]. We have shown that some strains identified by Spoligotyping as T, U or even orphan, which represent in our study the 52.