Inactivation du mycobacteriophage D29 par le sulfate ferreux d'ammonium pour la detection de Mycobacterium smegmatis et de M. tuberculosis viables

Add time:09/26/2019    Source:infona.pl

There is still an urgent requirement for more sensitive, cost-effective methods for detection and susceptibility testing of mycobacteria in clinical samples. We have been investigating a simple bacteriophage-based system which could be used for both purposes. As this depends upon the detection of phages which have successfully infected cells, a key step is the efficient removal or inactivation of phages remaining free in the culture medium. We demonstrate here the use of ferrous ammonium sulphate as an effective agent for the inactivation of mycobacteriophage D29 without impairing phage replication in previously infected host bacteria. Using this property, we report the detection of viable Mycobacterium smegmatis, M. bovis BCG and M. tuberculosis using simple low-cost technology. The method is highly sensitive, since it is able to detect 10 colony-forming units of M. smegmatis. It is also rapid, with the detection of M. tuberculosis in sputum specimens within 48 h.Nous avons mis en oeuvre une methode bacteriophagique simple pour la detection des mycobacteries dans des prelevements cliniques et pour l'evaluation de leur sensibilite aux antibiotiques. Cette methode etant basee sur le depistage des phages qui ont infecte les bacteries, l'etape determinante consiste a eliminer efficacement ou a inactiver les phages restes libres dans le milieu de culture. Le sulfate ferreux d'ammonium se montre efficace pour l'inactivation du mycobacteriophage D29 sans alteration de la replication intrabacterienne, ce qui nous a permis de deceler des cellules viables de Mycobacterium smegmatis, de M. bovis BCG et de M. tuberculosis d'une maniere simple et peu couteuse. Cette methode est tres sensible: elle peut detecter 10 unites formatrices de plages de M. smegmatis; et elle est rapide, pouvant deceler M. tuberculosis en 48 heures dans les expectorations.

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