JPS5818066B2 - Save Tsugaku Tekimetsukin Kakuninshaku - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a biological sterilization confirmation reagent, and its purpose is to be able to easily determine whether sterilization by gas sterilization, etc. has been completed. We aim to provide biological sterility confirmation reagents.

In recent years, various equipment and textile products, especially for hospitals, etc.

Gas sterilization such as ethylene oxide is now frequently used to sterilize plastic and rubber products.

However, it is not always easy to determine whether the sterilization has been completed completely, and therefore, conventional methods have been used to confirm this by using Bacillus subtilis, which is the most resistant to ethylene oxide gas.
s (globi gii), Bacillus s
By adsorbing a certain amount of spores of tea-rothermophilus etc. onto paperboard and putting it into a sterilizer together with the above-mentioned items to be sterilized, sterilization is performed, and after the sterilization, the spores are taken out and cultured for a certain period of time. , a method of biologically determining sterilization effectiveness has been adopted.

However, conventional test strips only adsorb the spores, so in order to confirm the survival of the spores, the spores on the sterilized test strip must be transferred to a designated medium and cultured to make a determination. However, since this requires special equipment, a furnace type, and skilled operation, this must be entrusted to a specialized organization capable of such operations.

However, since this takes a long time, it is extremely disadvantageous when an emergency is required due to its nature.

Therefore, the present inventor conducted various studies to solve these shortcomings, and as a result, the inventors of the present invention have developed an organism in which the effectiveness of sterilization can be easily determined by preparing a set of the above-mentioned spores and a medium used for this culture. We succeeded in obtaining a chemical sterility confirmation reagent.

That is, the present invention provides a biological sterilization confirmation reagent in which the spores of a test bacterium and a liquid medium are adsorbed to separate adsorbents and then dried to form a set, or both are adsorbed to the same adsorbent and then dried; Furthermore, it relates to a biological sterilization confirmation reagent sealed in an ampoule.

The test bacteria used in the present invention include bacteria resistant to ethylene oxide gas, such as Bacillus.
5ubtilis var, niger (CampDe
trick 5train), B, 5ubtilis
nigerS67, B, 5ubtilis var, g
lobigii, B.

pumilus (ATCC7061), B, aero
ther-mophi lus, B, cereus
, B. stearothermo-philus (
ATCC7953), Clostidium sporo
genes (ATCC7955), CJ? , 5p
oro-genes (ATCC3584), but especially B, 5ubtilis var, n
Preferably, iger is used.

These bacteria are grown in a medium where spores are easily formed, such as 4 g of potato extract, 4 g of yeast extract, and 2.5 g of glucose.
The spores are cultured in a conventional manner using a medium consisting of 15 g of agar, 15 g of agar, and 11 g of distilled water, and the spores are separated by a conventional method and added to distilled water to prepare a spore suspension.

Any liquid medium can be used as long as it can sufficiently culture the test prisoners, such as ordinary bouillon medium, potato medium, thioglycollate medium, Bart infusion medium, plain Bart infusion medium, and the like.

The spore suspension and liquid medium prepared as described above are adsorbed onto an adsorbent.

As the adsorbent, for example, F paper, cotton cloth, glass beads, ceramic tiles, etc. are used.

The spores 1 and the medium 2 may be adsorbed to separate adsorbents 3 as shown in FIG. 1 to form a set, or the spores 1 and the medium 2 may be adsorbed to the same adsorbent 3 as shown in FIG. You can also do that.

In the spore adsorption method, an adsorbent may be immersed in the spore suspension, but it is preferable to drop the spore suspension onto the adsorbent in order to adsorb a certain number of spores.

There are no particular limitations on the method of adsorbing the medium, and methods such as immersing the adsorbent in a liquid medium, dropping or spraying the liquid medium onto the adsorbent, and the like are employed.

The number of spores to be adsorbed to the adsorbent is not particularly limited, and the medium may be used in an amount that can sufficiently culture the spores, but for example, it is preferable to adsorb 10' to 10 6 spores and 100 to 500 spores in the medium.

The adsorbent with the spores and medium adsorbed in this manner is dried by air drying, hot air drying at a temperature that does not destroy the spores or medium components, vacuum drying, freeze drying, or the like.

The adsorbent obtained as described above is either aseptically packaged as it is, or sealed and stored in an airtight container such as an ampoule 4 as shown in FIG.

Since the biological sterilization confirmation reagent of the present invention is configured as described above, in order to use it to determine the sterilization effect, place the reagent together with the object to be sterilized in a sterilizer, perform sterilization work, After sterilization, the spores and culture medium are taken out. If the spores and the culture medium are adsorbed to separate adsorbents, they are combined as a set, and if both are adsorbed to the same adsorbent, they are sterilized and distilled in a predetermined amount. The spores and medium are eluted by placing in water, and the survival of the spores is determined by culturing at 37°C for 24 hours according to a conventional method.

Furthermore, if the reagent is sealed in an ampoule or the like, it is extremely convenient because a predetermined amount of sterile distilled water can be added to the ampoule and cultured in the same manner as described above.

As described above, since the biological sterilization confirmation reagent of the present invention has spores and a medium prepared at the same time, the sterilization effect can be determined by simple operations without requesting a specialized institution. Even if germs are present in the culture medium, they will be killed during the sterilization process, so the method has advantages such as accurate determination.

Next, an example will be given and explained.

; Example (1) Bacillus 5ubtilis va
An aqueous suspension of 106 gJ/ml of S. r.

A 15mm x 40mm Toyo Oki paper sheet 24 was sterilized by dry heat.
A water suspension of spores is placed 10 mm from one end of this filter paper.
．． Then, drop 1 ml of the above liquid medium into the remaining 30 mm and air dry.

This is sealed in an ampoule and stored.

(2) The results of determining the sterilization effect using the reagent prepared as described in (1) above are as follows.

Open the ampoule obtained in (1), put it in a gas sterilization chamber; put a lid on it, and add ethylene oxide powder 00 ~ /
l! ,, 54. ．． Sterilization was performed at 5° C. and 50% RH for a predetermined period of time.

After sterilization, remove the ampoule and add 5 m of sterile distilled water to it.
1 to crush the test paper, and shake the ampoule gently to dissolve the medium in the water.

This was covered with a sterilized cap and cultured at 37°C for 24 hours, and the cloudy state of the liquid was observed.

The results are as follows. As is clear from the results, it was confirmed that when the sterilization time was 10 minutes, the spores remained alive and the sterilization was incomplete, and when the sterilization time was 120 minutes, the spores were dead and the sterilization was complete.

[Brief explanation of the drawing]

Figure 1 shows the biological sterilization confirmation reagent when spores and culture medium are adsorbed on separate adsorbents, Figure 2 shows the biological sterilization confirmation reagent when spores and culture medium are adsorbed on the same adsorbent, and Figure 3 shows the biological sterilization confirmation reagent when spores and culture medium are adsorbed on the same adsorbent. This is a biological sterilization confirmation reagent in which an adsorbent in which spores and a culture medium are adsorbed is sealed in an airtight container. 1...Spore, 2...Medium, 3...
...Adsorbent, 4... Ampoule.

Claims (1)

[Scope of Claims] 1. Biology characterized by adsorbing spores of test bacteria and a liquid medium onto separate adsorbents and drying them to form a set, or adsorbing both onto the same adsorbent and drying them. Reagent for confirming sterility. 2. An organism characterized by adsorbing the spores of a test bacterium and a liquid medium onto separate adsorbents and then drying them to form a set, or by adsorbing both onto the same adsorbent, drying them, and sealing them in an airtight container. Scientific sterilization confirmation reagent 0