Pyrosequencing identification of Mycobacterium tuberculosis W-Beijing
The worldwide expanding Mycobacterium tuberculosis W-Beijing family is associated with treatment failure and relapse. Its identification currently relies on spoligotyping and conventional sequencing.
We developed pyrosequencing as an alternative method for its identification.FindingsPyrosequencing found a G/A substitution in the Rv0927c-pstS3 intergenic spacer and a RD105 deletion, identifying 8/104 M. tuberculosis isolates as W-Beijing isolates.
In addition, pyrosequencing found a previously unreported TGC deletion in the Rv0927c gene of W-Beijing isolates. Total concordance was found between the pyrosequencing data and conventional sequencing, as well as reference molecular identification.
Multispacer Sequence Typing assigned the W-Beijing isolates to the Asian lineage and the 96 non-W-Beijing isolates to the Euro-American lineage (P inferior 10 to 5). The W-Beijing isolates were all susceptible to streptomycin, rifampin, isoniazid, ethambutol, and pyrazinamide; no resistance-associated mutations were detected in these eight W-Beijing isolates.
There were no statistically significant differences in the antibiotic susceptibility of W-Beijing and non-W-Beijing isolates (p=0.2, X chi -2 test). Pyrosequencing correctly identified M.tuberculosis organisms in 26/26 sputum specimens exhibiting acid-fact bacilli.
Pyrosequencing results were obtained within four hours, incurring an estimated cost of 1.86 Euro/test.
Conclusions: Pyrosequencing of the Rv0927c gene and adjacent intergenic spacer is an efficient, low-cost technique for the rapid identification of W-Beijing isolates.
Author: Zoheira DjelouadjiMireille HenryAmine BachtarziNadege FoiselleDidier RaoultMichel Drancourt
Credits/Source: BMC Research Notes 2009, 2:239
Published on: 2009-12-02
Copyright by the authors listed above - made available via BioMedCentral (Open Access). Please
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