阅读说明：本技术 一种用于血培养瓶生产的试剂及血培养瓶生产工艺 (Reagent for blood culture bottle production and blood culture bottle production process ) 是由 孙月鹏 许琼杰 张娟丽 吕斌 谭韦丽 罗江卫 于 2019-10-22 设计创作，主要内容包括：本发明涉及医学化学发光免疫分析检测技术领域,具体涉及是一种用于血培养瓶生产的培养基及血培养瓶生产工艺,包括第一试剂制备、第二试剂制备和灌装入瓶。其中第一试剂包括硅胶RT601A、硅胶RT601B和氢氧化钠,第二试剂包括胰酪胨、脑心浸粉、明胶胨、葡萄糖、氯化钠、L-精氨酸、丙酮酸钠、SPS和Tris,通过设置在培养瓶底部的CO<Sub>2</Sub>传感器对样本活性进行检测,解决了现有血培养瓶基于传统检测方法检出率不高,且结果不稳定,采用新的检测方法未有与之相适配血培养瓶的问题。(The invention relates to the technical field of medical chemiluminescence immunoassay detection, in particular to a culture medium for blood culture bottle production and a blood culture bottle production process. Wherein the first reagent comprises silica gel RT601A, silica gel RT601B and sodium hydroxide, the second reagent comprises tryptone, brain heart extract powder, gelatin peptone, glucose, sodium chloride, L-arginine, sodium pyruvate, SPS and Tris, and the first reagent is prepared by placing CO at the bottom of a culture bottle 2 The sensor detects the activity of the sample, solves the problems that the detection rate of the existing blood culture bottle based on the traditional detection method is not high, the result is unstable, and the new detection method is adopted and the blood culture bottle is not matched with the traditional detection methodThe problem of bottle cultivation.)

1. A culture medium for blood culture flask production, characterized in that: including first reagent and second reagent, first reagent includes silica gel RT601A, silica gel RT601B and sodium hydroxide, the second reagent includes pancreas casein, brain heart soaks powder, gelatin peptone, glucose, sodium chloride, L-arginine, sodium pyruvate, SPS and Tris.

2. A blood culture bottle production process is characterized in that: the method comprises the steps of preparing a first reagent, preparing a second reagent and filling the reagent into a bottle, wherein the preparation of the first reagent comprises the following steps:

s1, preparing a reagent, namely taking a proper amount of first reagent, and adding purified water into the first reagent to prepare a silica gel mixture;

s2, stirring, and uniformly stirring the silica gel mixture;

s3, the first filling,filling the uniformly stirred silica gel mixture until the bottom is filled with CO₂Sensor and permselective CO₂The membrane of (a);

s4, drying, heating the culture bottle to dry and solidify the silica gel mixture at the bottom of the culture bottle;

s5, sterilizing, namely sterilizing the culture bottle after drying and curing for later use;

the second reagent preparation comprises the following steps:

s1, preparing a reagent, namely taking a proper amount of second reagent, and adding purified water into the second reagent to prepare a reagent mixed solution;

s2, carrying out damp-heat sterilization, namely adding the reagent mixed solution into a culture medium preparation device after uniformly stirring, and carrying out sterilization in a damp-heat environment;

the bottling comprises the following steps:

s1, filling for the second time, namely taking the sterilized culture bottles prepared in the S5 prepared by the first reagent and the sterilized culture medium prepared in the S2 prepared by the second reagent, and filling the sterilized culture medium into the sterilized culture bottles;

s2, plugging and capping, and sealing the capped culture bottle;

s3, labeling, wherein a label is attached to the surface of the culture bottle after the capping.

3. A blood culture bottle production process according to claim 2, wherein: in the first reagent preparation step S1, the silica gel RT601A is 800-1000 parts by weight, the silica gel RT601B is 50-150 parts by weight, the sodium hydroxide is a sodium hydroxide solution with the concentration of 1mol/L, the addition amount is 2-4 parts by weight, and the addition amount of the purified water is 20-40 parts by weight.

4. A blood culture bottle production process according to claim 3, wherein: in the first reagent preparing step S1, the silica gel RT601A is 900 parts by weight, the silica gel RT601B is 100 parts by weight, the sodium hydroxide is a sodium hydroxide solution having a concentration of 1mol/L, the addition amount is 3 parts by weight, and the addition amount of purified water is 30 parts by weight.

5. A blood culture bottle production process according to claim 4, wherein: in the second reagent preparation step S1, 13-17 parts of tryptone, 7-9 parts of brain heart extract powder, 8-12 parts of gelatin peptone, 3-7 parts of glucose, 3-7 parts of sodium chloride, 0.5-1.5 parts of L-arginine, 2-4 parts of sodium pyruvate, 0.15-0.35 part of SPS, 1.4-1.8 parts of Tris and 850-1150 parts of purified water are calculated by weight.

6. A blood culture bottle production process according to claim 5, wherein: in the second reagent preparation step S1, 15 parts of tryptone, 8 parts of brain-heart extract powder, 10 parts of gelatin peptone, 5 parts of glucose, 5 parts of sodium chloride, 1 part of L-arginine, 3 parts of sodium pyruvate, 0.25 part of SPS and 1.6 parts of Tris are calculated by weight, and the amount of the purified water added is 1000 parts.

7. A blood culture bottle production process according to claim 6, wherein: in the second reagent preparation step S2, the mixture is heated at 120 to 130 ℃ for 30min to perform moist heat sterilization.

8. A blood culture bottle production process according to claim 2, wherein: after the step of filling into the bottle, sampling detection is carried out, and the sampling detection comprises the following steps:

s1, extracting a certain amount of culture medium from each batch in the label-pasted culture bottle, and checking whether the culture medium is qualified or not;

and S2, warehousing the batches meeting the qualification rate, and destroying the batches not meeting the qualification rate.

Technical Field

The invention relates to the technical field of medical chemiluminescence immunoassay detection, in particular to a culture medium for blood culture bottle production and a blood culture bottle production process.

Background

Bacteremia or fungemia develops when microorganisms invade the blood and multiply rapidly beyond the body's immune system's ability to eliminate these microorganisms, and can infect extravascular tissues. The common method for detecting the presence of microorganisms in blood in clinical laboratories is blood culture, which is to collect a sample of patient blood and inoculate the sample into a culture flask for the discovery and identification of pathogenic microorganisms. The culture of living microorganisms in the blood circulation of patients has very important significance for the diagnosis and prognosis of patients. The positive blood culture results not only establish but also confirm that the disease of the patient is caused by infection with pathogenic bacteria, and more importantly, it also provides a test of the susceptibility of the pathogenic bacteria to antibiotics, thereby optimizing antibiotic therapy. From a prognostic perspective, the growth of a clinically important pathogen in blood culture indicates a decrease in resistance at the primary site of infection of the host, or the inability of the body to clear the pathogen or eradicate the focus of infection. The species of pathogenic bacteria cultured from blood are also of great significance for prognosis.

Chinese patent publication No. CN104450506A discloses an anaerobic blood culture medium, a culture flask and a preparation method of the culture flask, wherein two culture medium components with different compositions are mainly relied on, although cytotoxic substances generated in the metabolic process of the culture medium can be reduced, the traditional detection method is relied on, and the overall detection rate is still not high.

Disclosure of Invention

The invention provides a culture medium for blood culture bottle production and a blood culture bottle production process, aiming at the problems that the detection rate of the existing blood culture bottle based on the traditional detection method is not high, the result is unstable, and a new detection method is adopted and a blood culture bottle is not matched with the existing blood culture bottle.

The invention solves the technical problems and adopts the technical scheme that the culture medium for producing the blood culture bottle comprises a first reagent and a second reagent, wherein the first reagent comprises silica gel RT601A, silica gel RT601B and sodium hydroxide, and the second reagent comprises tryptone, brain heart soaking powder, gelatin peptone, glucose, sodium chloride, L-arginine, sodium pyruvate, SPS and Tris.

Further, the application also provides a blood culture bottle production process, which comprises the steps of preparing a first reagent, preparing a second reagent and filling the reagent into a bottle, wherein the preparation of the first reagent comprises the following steps:

s1, preparing a reagent, namely taking a proper amount of first reagent, and adding purified water into the first reagent to prepare a silica gel mixture;

s2, stirring, and uniformly stirring the silica gel mixture;

s3, filling the first time, namely filling the uniformly stirred silica gel mixture until the bottom is filled with CO₂Sensor and permselective CO₂The membrane of (a);

s4, drying, heating the culture bottle to dry and solidify the silica gel mixture at the bottom of the culture bottle;

s5, sterilizing, namely sterilizing the culture bottle after drying and curing for later use;

the preparation of the second reagent comprises the following steps:

s1, preparing a reagent, namely taking a proper amount of second reagent, and adding purified water into the second reagent to prepare a reagent mixed solution;

s2, carrying out damp-heat sterilization, namely adding the reagent mixed solution into a culture medium preparation device after uniformly stirring, and carrying out sterilization in a damp-heat environment;

the bottling comprises the following steps:

s1, filling for the second time, namely taking the sterilized culture bottles prepared in the S5 prepared by the first reagent and the sterilized culture medium prepared in the S2 prepared by the second reagent, and filling the sterilized culture medium into the sterilized culture bottles;

s2, plugging and capping, and sealing the capped culture bottle;

s3, labeling, wherein a label is attached to the surface of the culture bottle after the capping.

Further, in the first reagent preparing step S1, the silica gel RT601A is 800 to 1000 parts by weight, the silica gel RT601B is 50 to 150 parts by weight, the sodium hydroxide is a sodium hydroxide solution with a concentration of 1mol/L, the addition amount is 2 to 4 parts by weight, and the addition amount of the purified water is 50 to 100 parts by weight.

Optionally, in the first reagent preparing step S1, the silica gel RT601A is 900 parts by weight, the silica gel RT601B is 100 parts by weight, the sodium hydroxide is a sodium hydroxide solution with a concentration of 1mol/L, the addition amount is 3 parts by weight, and the addition amount of the purified water is 70 parts by weight.

Further, in the second reagent preparation step S1, 13-17 parts of tryptone, 7-9 parts of brain heart infusion powder, 8-12 parts of gelatin peptone, 3-7 parts of glucose, 3-7 parts of sodium chloride, 0.5-1.5 parts of L-arginine, 2-4 parts of sodium pyruvate, 0.15-0.35 part of SPS, 1.4-1.8 parts of Tris and 850-1150 parts of purified water are calculated by weight.

Optionally, in the second reagent preparing step S1, the weight parts of tryptone, brain heart extract powder 8, gelatin peptone 10, glucose 5, sodium chloride 5, L-arginine 1, sodium pyruvate 3, SPS 0.25, Tris 1.6, and purified water 1000.

Optionally, in the second reagent preparing step S2, heating is performed at 120 to 130 ℃ for 30min to perform wet heat sterilization.

Further, after the step of filling and bottling, sampling detection is also carried out, and the sampling detection comprises the following steps:

s1, extracting a certain amount of culture medium from each batch in the label-pasted culture bottle, and checking whether the culture medium is qualified or not;

and S2, warehousing the batches meeting the qualification rate, and destroying the batches not meeting the qualification rate.

The beneficial effects of the invention at least comprise one of the following;

1. the first reagent consisting of silica gel RT601A, silica gel RT601B and sodium hydroxide and the second reagent consisting of tryptose, casein, brain heart extract powder, gelatin peptone and the like are selected, wherein the first reagent mainly provides necessary matrix for the whole culture medium, and the second reagent provides necessary nutrient components for the culture medium, so that the culture is convenientThe cultured sample produces sufficient CO₂Growth in culture medium to produce CO, if viable microorganisms are present, after inoculation of the sample into a blood culture flask₂. CO through the bottom of the blood culture bottle₂Sensor, CO₂Sensor for CO permeation by selective₂Is separated from the liquid medium, CO₂Hydrogen ions can be displaced through the membrane and diffuse, acidifying the sensor, causing a color change in the sensor and causing a change in the signal of the reflected light on the monitoring system. The instrument judges whether microorganisms grow in the culture bottle by monitoring the change of the optical signal of the sensor, and compared with the traditional detection method, the detection rate is high, and the result is stable.

2. Solves the problems that the prior blood culture bottle has low detectable rate and unstable result based on the traditional detection method, and a new detection method is adopted and a blood culture bottle matched with the prior detection method is not available.

Drawings

FIG. 1 is a flow chart of a blood culture bottle production process.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the scope of the invention.