Novel gonococcus preservative solution

Abstract

The invention relates to the technical field of microorganism preservation, in particular to a novel gonococcus preservation solution which comprises the following components in parts by mass: 1-2 parts of a preservation agent, 40-60 parts of a solvent and 15-25 parts of a protective agent. According to the novel gonococcal preservation solution provided by the invention, the 30% glyceropancreatin soybean broth has a better gonococcal preservation effect than the conventional 10% skim milk at-20 DEG C, so that the gonococcal preservation of outpatient service and basic medical institutions with limited laboratory conditions is facilitated.

Description

Technical Field

[0001] The invention relates to the technical field of microbial preservation, in particular to a novel gonococcal preservation solution. Background Art

[0002] Gonorrhea is a notifiable Class B infectious disease in my country and one of the sexually transmitted diseases currently under priority prevention and control. The pathogen, gonococci, is a fastidious bacterium with high nutritional requirements, making cultivation and preservation difficult. Long-term preservation of gonococci presents numerous challenges. Currently, laboratories commonly use skim milk to preserve gonococci, but this method suffers from unstable preservation and short survival times, creating significant challenges for researchers involved in clinical testing, drug resistance monitoring, and related research.

[0003] Although there are various methods for preserving bacterial strains, such as solid culture medium, semi-solid puncture, drying, and cryopreservation, ideal bacterial preservation must not only maintain the activity of the strain but also ensure its inherent biological properties and antigenic properties. Gonorrhea has weak resistance to external physical and chemical factors, is sensitive to heat, and is prone to producing autolytic enzymes. Conventional solid culture media require regular subculture, which is cumbersome and prone to strain mutation. Although freeze-drying is considered the optimal method for bacterial preservation, it is complex, time-consuming, labor-intensive, and requires expensive equipment. Conventional 10% skim milk for preserving gonococci has poor preservation performance, maintaining them for only one day at 4°C and for four days at -18°C and -29°C, without changing their bacterial count and activity. For primary healthcare institutions with limited laboratory resources, there is an urgent need for a cost-effective, stable, and reliable method for preserving gonococci. Summary of the Invention

[0004] The object of the present invention is to provide a novel gonococcal preservation solution to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a new type of gonococcal preservation solution, comprising the following components by mass: 1-2 parts of preservative, 40-60 parts of solvent, and 15-25 parts of protective agent.

[0006] Preferably, the storage temperature of the gonococcal preservation solution is -20°C or -70°C.

[0007] Preferably, the preservative is tryptic soy broth.

[0008] Preferably, the solvent is distilled water.

[0009] Preferably, the protective agent is glycerol.

[0010] Compared with the prior art, the present invention has the following beneficial effects:

[0011] The invention proposes a novel gonococcal preservation solution, 30% glycerol trypsin soy broth, which has a better effect on preserving gonococci at -20°C than conventional 10% skim milk, and is beneficial for preserving gonococci in outpatient clinics and primary medical institutions with limited laboratory conditions. DETAILED DESCRIPTION

[0012] The technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0013] The present invention provides a technical solution: a novel gonococcal preservation solution, the storage temperature of the gonococcal preservation solution is -20°C or -70°C, and the gonococcal preservation solution comprises the following components by mass: 1-2 parts of a preservative, which is trypsin soy broth, 40-60 parts of a solvent, which is distilled water, and 15-25 parts of a protective agent, which is glycerol.

[0014] The steps for preparing the gonococcal preservation solution are as follows:

[0015] 1. Weigh 1-2g of tryptic soy broth, add 40-60ml of distilled water, mix well and autoclave at 121°C for 15 minutes.

[0016] 2. Measure 15-25 ml of glycerol and autoclave at 116°C for 5 minutes.

[0017] 3. Take out the sterilized 3% trypsin soy broth, add it to glycerol, mix well, divide into portions, and place in a -20℃ or -70℃ refrigerator for later use.

[0018] Specific embodiment (preparing 60 ml of 30% glycerol trypsin soy broth as an example): weigh 1.5 ml of trypsin soy broth, add 50 ml of distilled water, mix well, and then autoclave, the autoclaving temperature and time are 121°C and 15 minutes; measure 18 ml of glycerol, and autoclave the temperature and time at 116°C and 5 minutes; take out 42 ml of the sterilized 3% trypsin soy broth, add it to the sterilized 30% glycerol, mix well, divide into portions, and place in a -20°C or -70°C refrigerator for use.

[0019] In order to verify the difference between the preservation effect of 30% glycerol trypsin soy broth and conventional 10% skim milk in the preservation of gonococci, several comparative experiments were conducted. Among them, the effect of 10% skim milk in the preservation of gonococci at different temperatures is shown in the following table:

[0020] Observation time 4℃ -18℃ -29℃ -70℃ liquid nitrogen same day ++++ ++++ ++++ ++++ ++++ 1 day ++↑ ++++ ++++ ++++ ++++ 2 days ++↓ ++++ ++++ ++++ ++++ 3 days 3 ++++ ++++ ++++ ++++ 4 days — ++++ ++++ ++++ ++++ 1 week ++↑ +++ ++++ ++++ 2 weeks ++↑ +++ ++++ ++++ 3 weeks ++ +++ ++++ ++++ 1 month 10 12 ++++ ++++ 2 months — — ++++ ++++ 3 months ++++ ++++ 6 months ++++ ++++

[0021] In addition to 30% glycerol trypsin soy broth, three other preservation solutions were also made. The following table shows the effects of these four preservation solutions on the preservation of gonococci at -20°C:

[0022] Observation time 10% DMSO calf serum 10% glycerol calf serum 20% glycerol brain heart infusion 30% glycerol tryptic soy broth 1 week +++ ++++ ++++ ++++ 2 weeks Some survive +++ ++++ ++++ 1 month A few survive Some survive ++++ ++++ 2 months - - ++++ ++++ 3 months - - ++++ ++++ 6 months - - - A few survive 9 months - - - - 12 months - - - -

[0023] Therefore, the 30% glycerol trypsin soy broth proposed by the present invention has a better effect on preserving gonococci at -20°C than conventional 10% skim milk, and is beneficial for preserving gonococci in outpatient clinics and primary medical institutions with limited laboratory conditions.

[0024] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the underlying principles of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A novel gonococcal preservation solution, characterized in that: The invention comprises the following components by weight: 1-2 parts of preservative, 40-60 parts of solvent and 15-25 parts of protective agent.

2. A novel gonococcal preservation solution according to claim 1, characterized in that: The storage temperature of the gonococcal preservation solution is -20°C or -70°C.

3. A novel gonococcal preservation solution according to claim 1, characterized in that: The preservative is tryptic soy broth.

4. A novel gonococcal preservation solution according to claim 1, characterized in that: The solvent is distilled water.

5. A novel gonococcal preservation solution according to claim 1, characterized in that: The protective agent is glycerol.