US 12,448,642 B2
Biological indicator for determining the efficacy of an oxidative sterilization process and methods of use
Adrián Jesús Rovetto, Alvear (AR); Esteban Lombardía, Alvear (AR); Pablo Ravasi, Alvear (AR); and Carlos Ernesto Rodriguez, Alvear (AR)
Assigned to TERRAGENE LLC, Spring, TX (US)
Filed by TERRAGENE LLC, Spring, TX (US)
Filed on Jul. 2, 2021, as Appl. No. 17/367,224.
Prior Publication US 2023/0002802 A1, Jan. 5, 2023
Int. Cl. C12Q 1/22 (2006.01); G01N 21/64 (2006.01)
CPC C12Q 1/22 (2013.01) [G01N 21/6486 (2013.01)] 19 Claims
 
1. A biological indicator for determining the efficacy of an oxidative sterilization process, wherein said biological indicator comprises, in a single container,
a set of microbial spores,
at least one fluorescent sensor protein, and
a culture medium,
wherein the at least one fluorescent sensor protein is capable of yielding an optically detectable signal when the at least one fluorescent sensor protein is not in a denatured state due to the oxidative sterilization process, the at least one fluorescent protein being further capable of yielding a different optically detectable signal when the at least one sensor protein is in a denatured state after the oxidative sterilization process, said difference being independent of a catalytic activity of the at least one fluorescent sensor protein and the culture medium being brought into contact with the set of microbial spores,
wherein said culture medium is capable of inducing growth of any viable microbial life present after the oxidative sterilization process,
wherein said culture medium comprises a colorimetric component being capable of undergoing an optically detectable color shift in the presence of microbial growth,
wherein denaturation of the fluorescent sensor protein is correlated with death of the microbial spores, a decrease in the optically detectable signal being indicative of a successful sterilization process, and
wherein said fluorescent sensor protein is exogenous from said microbial spores and said decrease is detectable instantaneously after the sterilization process.