Method for isolating and culturing unculturable microorganisms

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 1/20 (2006.01) C12N 1/00 (2006.01) C12N 9/80 (2006.01) C12P 21/04 (2006.01) C12Q 1/02 (2006.01)

Patent

CA 2498243

The invention provides a method for isolating and culturing a previously unculturable microorganism, which comprises: (i) collecting a sample from an environmental source; (ii) counting/estimating the number of microorganisms in the sample; (iii) diluting the sample in an appropriate medium; (iv) adding a gelating agent such as to entrap one or more microorganisms within a sphere of the gelating agent; (v) coating the spheres containing the entrapped microorganism(s) with a natural or synthetic polymer to form a polymeric membrane; (vi) incubating the coated spheres in the original environment for an appropriate time; (vii) cutting the spheres and scanning for microorganisms colonies; and (viii) isolating the microorganisms, and repeating steps (iii) to (vii) until a pure clone of said previously unculturable microorganism is obtained.

L'invention concerne un procédé permettant d'isoler et de faire croître en milieu de culture un micro-organisme qui ne pouvait auparavant pas croître en milieu de culture. Ce procédé consiste à : (i) prélever un échantillon d'une source environnementale ; (ii) compter/estimer le nombre de micro-organismes contenus dans cet échantillon ; (iii) diluer l'échantillon dans un milieu approprié ; (iv) ajouter un agent gélifiant pour piéger un ou plusieurs micro-organisme(s) dans une sphère de cet agent gélifiant ; (v) appliquer un polymère naturel ou synthétique sur les sphères contenant le(s) micro-organisme(s) piégé(s) pour former une membrane polymère ; (vi) incuber les sphères ainsi revêtues dans l'environnement d'origine pendant une durée appropriée ; (vii) couper ces sphères et les examiner pour détecter des colonies de micro-organismes ; et (viii) isoler lesdits micro-organismes. Les étapes (iii) à (vii) sont répétées jusqu'à l'obtention d'un clone pur du micro-organisme qui ne pouvait auparavant pas croître en milieu de culture.

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