1887

Abstract

Isolated more than half a century ago, the H37Rv strain of still remains the strain of choice for the majority of laboratories conducting studies of TB pathogenesis. In this report we reveal that H37Rv is highly prone to losing the ability to synthesize the cell wall lipid phthiocerol dimycocerosate (PDIM) during extended periods of culture. In addition, H37Rv stocks that have been held for even a short length of time should be thought of as a heterogeneous population of PDIM-positive and PDIM-negative cell types. We demonstrate that after weekly subculture of PDIM-positive isolates over a period of 20 weeks, the proportion of PDIM-negative cells rises above 30 %. That PDIM biosynthesis is negatively selected is evident from the broad range of mutation types we observe within cultures originating from a single PDIM-positive parental clone. Moreover, the appearance of these multiple mutation types coupled with an enhanced growth rate of PDIM-negative bacteria ensures that ‘PDIM-less’ clones rapidly dominate cultures. It has been known for almost a decade that strains of that lack PDIM are severely attenuated during infection. Therefore, the loss of PDIM raises a very serious issue in regard to the interpretation of putative virulence factors where heterogeneous parental cultures are potentially being compared to recombinant clones isolated within a PDIM-negative background. It is essential that researchers undertaking virulence studies confirm the presence of PDIM within all recombinant clones and the parental strains they are derived from.

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2009-11-01
2019-10-18
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