PMID- 11283094
OWN - NLM
STAT- MEDLINE
DCOM- 20010823
LR  - 20181113
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 39
IP  - 4
DP  - 2001 Apr
TI  - Use of spoligotyping to study the evolution of the direct repeat locus by IS6110 
      transposition in Mycobacterium tuberculosis.
PG  - 1595-9
AB  - Based on the variability of 43 spacers within the direct repeat (DR) locus of 
      Mycobacterium tuberculosis complex organisms, spoligotyping is a rapid method 
      that aids in the study of the epidemiology of tuberculosis. It was recently 
      hypothesized that despite its presence in the DR locus, spacer 31 could not be 
      amplified in M. tuberculosis clinical isolates belonging to spoligotype 50 due to 
      the insertion of an extra copy of IS6110 between spacers 31 and 32 that could 
      lead to an asymmetrical split of the primer targets (I. Filliol, C. Sola, and N. 
      Rastogi, J. Clin. Microbiol. 38:1231--1234, 2000). In the present investigation, 
      previous observations were extended to 25 clinical isolates of type 50 showing 
      that the primer set IS6-DRb that selectively amplified the left and central DR 
      regions was indeed able to demonstrate the presence of spacer 31. 
      IS6110-restriction fragment length polymorphism (RFLP) and DR-RFLP showed that 
      type 50 isolates were characterized by the presence of two copies of IS6110 
      associated with the DR locus and an additional double IS6110 band of 1.4 kb. The 
      primer set IS3-IS6 was then used to selectively amplify a 750-bp inter-IS6110 
      fragment within the DR locus. The sequencing of the central DR region 
      corroborated our previous findings and showed that the absence of spacer 31 among 
      the type 50 isolates was due to the asymmetric insertion of an extra copy of 
      IS6110 between spacers 31 and 32, leading to an unequal split of the DRa-DRb 
      target into two portions, of 6 and 30 bp, respectively. These results show that 
      the DR locus constitutes an ideal IS6110 preferential locus (ipl), permitting the 
      insertion of two or more copies of IS6110, and provide new clues for 
      epidemiological and phylogenetic interpretation of changes in IS6110-RFLP and 
      spoligotyping profiles.
FAU - Legrand, E
AU  - Legrand E
AD  - Unite de la Tuberculose et des Mycobacteries, Institut Pasteur de Guadeloupe, 
      97165 Pointe-a-Pitre Cedex, Guadeloupe.
FAU - Filliol, I
AU  - Filliol I
FAU - Sola, C
AU  - Sola C
FAU - Rastogi, N
AU  - Rastogi N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA Transposable Elements)
RN  - 0 (DNA, Bacterial)
RN  - 0 (DNA, Intergenic)
RN  - 0 (Oligodeoxyribonucleotides)
SB  - IM
MH  - Bacterial Typing Techniques
MH  - Base Sequence
MH  - *DNA Transposable Elements
MH  - DNA, Bacterial/genetics
MH  - DNA, Intergenic/*genetics
MH  - *Evolution, Molecular
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mycobacterium tuberculosis/classification/*genetics
MH  - Oligodeoxyribonucleotides/genetics
MH  - Polymerase Chain Reaction
MH  - Polymorphism, Restriction Fragment Length
MH  - Repetitive Sequences, Nucleic Acid/*genetics
MH  - Sequence Analysis, DNA
PMC - PMC87977
EDAT- 2001/04/03 10:00
MHDA- 2001/08/29 10:01
PMCR- 2001/04/01
CRDT- 2001/04/03 10:00
PHST- 2001/04/03 10:00 [pubmed]
PHST- 2001/08/29 10:01 [medline]
PHST- 2001/04/03 10:00 [entrez]
PHST- 2001/04/01 00:00 [pmc-release]
AID - 1318 [pii]
AID - 10.1128/JCM.39.4.1595-1599.2001 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2001 Apr;39(4):1595-9. doi: 10.1128/JCM.39.4.1595-1599.2001.