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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 Antimicrob+Agents+Chemother
2015 ; 59
(10
): 6521-38
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gab.com Text
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English Wikipedia
Rapid, Semiquantitative Assay To Discriminate among Compounds with Activity
against Replicating or Nonreplicating Mycobacterium tuberculosis
#MMPMID26239979
Gold B
; Roberts J
; Ling Y
; Quezada LL
; Glasheen J
; Ballinger E
; Somersan-Karakaya S
; Warrier T
; Warren JD
; Nathan C
Antimicrob Agents Chemother
2015[Oct]; 59
(10
): 6521-38
PMID26239979
show ga
The search for drugs that can kill replicating and nonreplicating Mycobacterium
tuberculosis faces practical bottlenecks. Measurement of CFU and discrimination
of bacteriostatic from bactericidal activity are costly in compounds, supplies,
labor, and time. Testing compounds against M. tuberculosis under conditions that
prevent the replication of M. tuberculosis often involves a second phase of the
test in which conditions are altered to permit the replication of bacteria that
survived the first phase. False-positive determinations of activity against
nonreplicating M. tuberculosis may arise from carryover of compounds from the
nonreplicating stage of the assay that act in the replicating stage. We mitigate
these problems by carrying out a 96-well microplate liquid MIC assay and then
transferring an aliquot of each well to a second set of plates in which each well
contains agar supplemented with activated charcoal. After 7 to 10 days-about 2
weeks sooner than required to count CFU-fluorometry reveals whether M.
tuberculosis bacilli in each well have replicated extensively enough to reduce a
resazurin dye added for the final hour. This charcoal agar resazurin assay (CARA)
distinguishes between bacterial biomasses in any two wells that differ by 2 to 3
log10 CFU. The CARA thus serves as a pretest and semiquantitative surrogate for
longer, more laborious, and expensive CFU-based assays, helps distinguish
bactericidal from bacteriostatic activity, and identifies compounds that are
active under replicating conditions, nonreplicating conditions, or both. Results
for 14 antimycobacterial compounds, including tuberculosis (TB) drugs, revealed
that PA-824 (pretomanid) and TMC207 (bedaquiline) are largely bacteriostatic.
free
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