BACKGROUND: Mycobacterial Interspersed Repetitive Units (MIRUs) that are located mainly in intergenic regions dispersed throughout the Mycobacterium tuberculosis genome. The selected MIRU loci, which were composed of a 12-locus set, demonstrated a high power for discrimination of Mycobacterium tuberculosis isolates collected from Kangwon province of Korea. To evaluate its ability to discriminate the M. tuberculosis strains, 45 clinical isolates were genotyped using the methods IS6110 RFLP and MIRU. METHODS: All the samples were collected during the period from January 2007 to December 2007 from TB patients, who were residents and registered to a public health center of Kangwon Province in Korea. A total of 45 DNAs were extracted from clinical isolated mycobacterial strains and genotyped using IS6110 RFLP, the MIRU method. RESULTS: We compared the 12-MIRU with IS6110 RFLP in the 45 samples, the 12-locus version offered less discriminatory power (Hunter-Gaston discriminatory index [HGDI]: 0.959 vs 0.998; 57.78% of clustered cases vs 8.89%). CONCLUSION: This 12-locus MIRU can be useful when additional combinations of other loci for genotyping M. tuberculosis in Korea where the Beijing family strains are dominant.
Discrimination (Psychology) ; DNA ; DNA, Intergenic ; Genome ; Genotype ; Humans ; Interspersed Repetitive Sequences ; Korea ; Mycobacterium ; Mycobacterium tuberculosis ; Polymorphism, Restriction Fragment Length ; Public Health ; Tuberculosis

We investigated the effect of alpha-subunit of the stimulatory GTP-binding protein (Galphas) gene mutation on the expression of the thyrotropin-releasing hormone (TRH) receptor (TRH-R) gene in GH3 cells and in growth hormone (GH)-secreting adenomas of acromegalic patients. In the presence of cycloheximide, forskolin and isobutylmethylxanthine, cholera toxin, and GH-releasing hormone (GBRH) decreased rat TRH-R (rTRH-R) gene expression by about 39%, 43.7%, and 46.7%, respectively. Transient expression of a vector expressing mutant-type Galphas decreased the rTRH-R gene expression by about 50% at 24 h of transfection, whereas a wild-type Galphas expression vector did not. The transcript of human TRH-R (hTRH-R) gene was detected in 6 of 8 (75%) tumors. Three of them (50%) showed the paradoxical GH response to TRH and the other three patients did not show the response. The relative expression of hTRH-R mRNA in the tumors from patients with the paradoxical response of GH to TRH did not differ from that in the tumors from patients without the paradoxical response. Direct PCR sequencing of GALPHAs gene disclosed a mutant allele and a normal allele only at codon 201 in 4 of 8 tumors. The paradoxical response to TRH was observed in 2 of 4 patients without the mutation, and 2 of 4 patients with the mutation. The hTRH-R gene expression of pituitary adenomas did not differ between the tumors without the mutation and those with mutation. The present study suggests that the expression of TRH-R gene is not likely to be a main determinant for the paradoxical response of GH to TRH, and that Galphas mutation may suppress the gene expression of TRH-R in GH-secreting adenoma. However, a certain predisposing factor(s) may play an important role in determining the expression of TRH-R.
Acromegaly ; Adenoma ; Alleles ; Animals ; Cholera Toxin ; Codon ; Colforsin ; Cycloheximide ; Gene Expression ; Growth Hormone ; GTP-Binding Proteins ; Humans ; Pituitary Neoplasms ; Polymerase Chain Reaction ; Rats ; Receptors, Thyrotropin-Releasing Hormone* ; RNA, Messenger* ; Thyrotropin-Releasing Hormone ; Transfection