BIOLOGICAL INDICATOR FOR RAPID VERIFICATION OF DISINFECTION OR STERILIZATION

Abstract

The present invention provides a biological indicator for verification of disinfection or sterilization, and in particular, provides a biological indicator capable of verifying disinfection or sterilization using luminescent microorganisms, and a kit for verification of disinfection or sterilization using the biological indicator. According to the present invention, it is possible to provide a biological indicator for verification of disinfection or sterilization that may confirm in real time how much disinfection or sterilization has been performed after disinfecting or sterilizing harmful microorganisms by irradiation with UV rays.

Claims

1. A biological indicator for rapid verification of disinfection or sterilization comprising a luminescent microorganism or a fragment of the luminescent microorganism.

2. The biological indicator for rapid verification of disinfection or sterilization according to claim 1, wherein the biological indicator verifies sterilization by irradiation with UV rays.

3. The biological indicator for rapid verification of disinfection or sterilization according to claim 1, wherein the luminescent microorganism is transformed by inserting a luciferase gene into a microorganism.

4. The biological indicator for rapid verification of disinfection or sterilization according to claim 3, wherein the luciferase gene is inserted before or after spore formation of the microorganism.

5. The biological indicator for rapid verification of disinfection or sterilization according to claim 3, wherein the luminescent microorganism is transformed by inserting a luciferase gene into any one or more strains selected from Salmonella enterica, Acinetobacter baumannii, and Porphyromonas gingivalis.

6. A kit for verification of disinfection or sterilization comprising a UV-ray transmittance cover and a spore carrier.

7. The kit for verification of disinfection or sterilization according to claim 6, wherein the spore carrier is inoculated with 10,000 cfu/ml to 1,000,000 cfu/ml of a luminescent microorganism, a spore type luminescent microorganism, or fragments thereof.

8. The kit for verification of disinfection or sterilization according to claim 6, wherein a material of the spore carrier is any one of stainless steel, cellulose paper, polystyrene, or polypropylene.

9. The kit for verification of disinfection or sterilization according to claim 6, wherein a material of the UV-ray transmittance cover is quartz.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0050] The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:

[0051] FIG. 1 is an enlarged view illustrating a configuration of a kit for verification of sterilization by irradiation with UV rays, which includes a biological indicator according to the present invention;

[0052] FIG. 2 is a plan view of the configuration shown in FIG. 1; and

[0053] FIG. 3 is a three-dimensional view of the configuration shown in FIG. 1.

DETAILED DESCRIPTION OF THE INVENTION

[0054] Hereinafter, the present invention will be described in detail by examples and experimental examples.

[0055] However, the following examples and experimental examples are only given for illustrating the present invention, and contents of the present invention are not limited to the following examples and experimental examples.

Example 1. Preparation of Biological Indicator

[0056] Intestinal microorganism Salmonella enterica was selected as a test strain, and Acinetobacter baumannii was selected as a respiratory microorganism, then a biological indicator was prepared by the following manner.

[0057] First, luciferase gene (pMCS-Red Firefly Luc Vector for Luciferase Assays (16155), ThermoFisher; pMCS-Gaussia Luc Vector for Luciferase Assays (16146), ThermoFisher) was subjected to ligation, followed by applying heat shock for transformation. After sequencing the vector, an activated cell (competent cell) was prepared so that DNA could be inserted. Thereafter, the cell was transformed using electroporation, followed by confirming whether the cell is transformed through colony-PCR.

Example 2. Preparation of Biological Indicator

[0058] The intestinal microorganism Salmonella enterica was selected as a test strain, and a biological indicator was prepared in the same manner as in Example 1.

Example 3. Preparation of a Biological Indicator

[0059] A biological indicator was prepared according to the same procedures as described in Example 1, except that oral microorganism Porphyromonas gingivalis was selected as a test strain.

Experimental Example 1. Confirmation of Sterilization Verification Effect by Measuring an Amount of Bioluminescence Expression of the Biological Indicator according to the Present Invention

[0060] An in-vitro experiment was performed to confirm a sterilization verification effect of the biological indicator including the intestinal microorganism and the respiratory microorganism prepared in Example 1. First, E. coli, and the like were smeared on a medium, and whether there is luminescence was measured before and after sterilization with UV-C having a wavelength of 100 to 280 nm. A luminescence measurement was performed through a primary observation using a simple inspection equipment utilizing a UV lamp (420 to 680 nm) in the field, and when clear quantitative evaluation was not possible in the primary observation, the luminescence was quantitatively measured using a charge-cooled digital (CCD) camera equipment. As a result of the experiment, it was immediately confirmed that, before sterilization with UV-C, luminescence of the biological indicator prepared in Example 1 was exhibited in a wide range, but after sterilization with UV-C, the luminescence range of the biological indicator was decreased.

Experimental Example 2. Confirmation of Sterilization Verification Effect by Measuring an Amount of Bioluminescence Expression of the Biological Indicator According to the Present Invention

[0061] An in-vivo experiment was performed to confirm a sterilization verification effect of the biological indicator including the intestinal microorganism prepared in Example 2. A mouse was prepared, and the biological indicator was administered into an abdomen of the mouse, then whether there is luminescence was measured measured before and after sterilization with UV-C having a wavelength of 100 to 280 nm. A luminescence measurement was performed through a primary observation using a simple inspection equipment utilizing a UV lamp (420 to 680 nm) in the field, and when clear quantitative evaluation was not possible in the primary observation, the luminescence was quantitatively measured using a charge-cooled digital (CCD) camera equipment. As a result of the experiment, it was immediately confirmed that, before sterilization with UV-C, luminescence of the biological indicator prepared in Example 2 was exhibited in a wide range, but after sterilization with UV-C, the luminescence range of the biological indicator was decreased.

Experimental Example 3. Confirmation of Sterilization Verification Effect by Measuring an Amount of Bioluminescence Expression of the Biological Indicator According to the Present Invention

[0062] An in-vitro experiment was performed to confirm a sterilization verification effect of the biological indicator including various types of oral microorganisms prepared in Example 3. First, a sample containing E. coli and the like and the biological indicator prepared in Example 3 were put together in a tube and sterilized with UV-C having a wavelength of 100 to 280 nm, and whether there is luminescence was measured before and after sterilization with UV-C having a wavelength of 100 to 280 nm. A luminescence measurement was performed through a primary observation using a simple inspection equipment utilizing a UV lamp (420 to 680 nm) in the field, and when clear quantitative evaluation was not possible in the primary observation, the luminescence was quantitatively measured using a charge-cooled digital (CCD) camera equipment. As a result of the experiment, it was immediately confirmed that, before sterilization with UV-C, luminescence of the biological indicator prepared in Example 3 was exhibited in a wide range, but after sterilization with UV-C, the luminescence range of the biological indicator was decreased.

[0063] The kit for verification of disinfection or sterilization, which includes the biological indicator according to the present invention may provide a reliable biological indicator while being capable of performing verification in real time. Further, the biological indicator and the kit may be used to accurately check quarantine results in real time, such that industrial applicability is very high.