Genotyping of the Mycobacterium tuberculosis complex using MIRUs: association with VNTR and spoligotyping for molecular epidemiology and evolutionary genetics.
The recent introduction of molecular methods has gained increasing acceptance as a powerful tool for epidemiology and phylogeny of tuberculosis (TB). In this study, the efficiency of using the molecular typing of mycobacterial interspersed repetitive unit (MIRUS) rated on a set of 116 clinical isolates of Mycobacterium tuberculosis complex of 11 different geographical origin. The result was compared with spoligotyping and variable number tandem repeat of DNA (VNTRs) data type. Eighty-nine Miru different profiles obtained in the studied sample. Spoligotyping- or VNTR cluster-defined, divided into subclusters by Miru typing.
In contrast, nearly all clinical isolates clustered by MIRUS shown belong to cluster-based spoligotyping defined. Calculation of discriminatory power by Hunter-Gaston index (HGI) for VNTR, spoligotyping and Miru typing gives values, respectively, 0.959, 0.965 and 0.988, indicating high discriminatory power of MIRUS. Allelic diversity samples are measured for each Miru-VNTR loci; five Miru locus (Miru nos. 10, 23, 26, 31 and 40) are “very discriminant”, four (Miru nos. 4, 16, 24 and 39) are “fairly discriminant”, and three (Miru nos. 2, 20 and 27) is “bad discriminant”. Among the three VNTRs complementary (exact tandem repeats ETR-A, ETR and ETR-B-C), ETR-A is the most discriminant locus. A combined numerical analysis of spoligotyping, VNTR typing and the results corroborated Miru most recent evolutionary scenario for M. tuberculosis complex hypothesis.
M. canettii will be the first branch that has deviated from the general M. tuberculosis complex ancestors. East-Africa India (EAI) clade could be the first family to have diverged thereafter. A third branching separating M. africanum-M. bovis clade, followed by a separate node versus non-Beijing Beijing M. tuberculosis. Beijing is a different clade of Central Asia 1 (CAS1) family. Among the non-Beijing strains, branches such as Latin America and the Mediterranean (LAM), X and Haarlem clades diverged later. In conclusion, the results obtained showed concordance between clades defined by spoligotyping, and Miru-VNTR, and underscores the potential of this method for M. tuberculosis phylogeny reconstruction. We also conclude that the Miru typing is a very promising method that can be used in genotyping strategy “PCR-based two”, in conjunction with conventional epidemiological investigation.
Genotyping of the Mycobacterium tuberculosis complex using MIRUs: association with VNTR and spoligotyping for molecular epidemiology and evolutionary genetics.
molecular epidemiological analysis of Cryptosporidium spp. in the UK: results of the genotype of Cryptosporidium spp. in 1705 from human stool samples and 105 samples of droppings from livestock.
Cryptosporidium present in stool samples of 1,705 people and 105 livestock were analyzed by PCR-restriction fragment length polymorphism Cryptosporidium oocyst wall protein. Overall, genotype 1 (exclusive type of man) was detected in 37.8% of samples from humans, and genotype 2 (broad host range) was detected in 61.5%, the third designated genotype genotype 3 (Cryptosporidium meleagridis) detected at 0, 3%, and both genotypes 1 and 2 have been recovered from 0.4%. All samples from cattle produce genotype 2.
Bovine Sex- determining region Y protein, SRY ELISA KIT
Among the 469 patients who were infected during the eight drinking water-related outbreaks, five outbreaks mainly occurred due to genotype 1, and three are due to genotype 2. Fifty-four samples collected from patients involved with five -associated plague swimming pool: two outbreaks are due to genotype 1, one is due to genotype 2, and the remaining two involved both genotype 1 and 2. among the 26 plague outbreak nurseries family and 1 children (2 to 3 members per group) , each genotype were recovered from different members of each outbreak: 13 is due to genotype 1, and 14 are due to genotype 2.
Kent
