SE448737B - PROCEDURE AND DEVICE FOR THE DISPOSAL OF BACTERIA ANIMALS - Google Patents

Description

448 737 the patient's vein through a cannula unit protected by DE-OS 2332133, a sterile syringe or a blood collection set- The withdrawn blood enters directly into a sterile vessel filled with liquid nutrient medium in the blood culture system, which with different volumes is designed as a glass tube, - bottle or vessel, closed with a rubber cap or metal lid. These known blood culture vessels are closed with sterile plugs; in the case of non-sterile clots, it is necessary that these be carefully decontaminated by means of a disinfectant before the blood draw.

By inserting the blood culture vessel into the holder of the cannula unit, the blood culture system is connected to the blood collection system. After puncturing the vein, the vessel is pressed against the bottom of the holder, so that the plug is pierced and thus immediately blood corresponding to the acting vacuum is sucked out of the vein. Through the blood, the liquid nutrient medium is immediately inoculated. If additional blood tests are required, several blood culture vessels are filled one after the other with different nutrient media.

Preparation and establishment of the subcultures: After the blood culture vessel has been removed from the holder of the cannula unit and thereby the blood collection system is separated from the blood culture system and after blood and liquid nutrient medium have been intensively mixed, the culture culture vessels are grown at 35-370 in preparation for the establishment of subcultures. The cultivation of aerobic and anaerobic microbes is carried out by fitting an unlocked or sealed aeration unit; after their growth, which is to be checked several times at fixed time intervals, after 448,737 any visible turbidity of the liquid nutrient medium, the subcultures of solid nutrient medium occur. At various time intervals, the cultured nutrient fluid mixed with microbes is removed from the blood culture vessel by means of a syringe via a sterile cannula, whereby the closure of the vessel is pierced. Inoculate one or more solid nutrient media, which are placed on separate plates or petri dishes and preferably consist of agar with different differentiated additives. After inoculation of the agar cultures, these are re-grown separately. the opening of the blood culture vessels for the blood inoculation takes place at fixed time intervals on the 3rd, 6th and 10th day after blood draw- Sh.

Only after the microbial growth in the subcultures is it possible to make unambiguous microbiological examinations of the various organisms.

The disadvantages of this known method are that the individual procedural steps for blood collection and establishment of blood cultures are performed with several devices, so that no safety is guaranteed for avoiding contamination during the draining and during construction and dismantling of the cultures.

For the blood culture systems operating according to this procedure, liquid culture medium vessels filled with glass with rubber or metal closures are known.

For the blood draw from the vein and for the blood inoculation as well as for the insufflation of gaseous substances in the culture system and for the inoculation of liquid nutrient media for the subcultures, it is constantly necessary to pierce resp. open these blood culture vessels. 448 737 In all these procedural steps and in particular in the subculture technology in the laboratory, contamination possibilities are given due to the drilling resp. the opening of the vessel closures and through the manipulation of syringes and needles.

A further disadvantage of this known method is that dilute bacterial colonies can hardly be distinguished either in liquid nutrient media or on solid agar.

According to further prospectus material, blood culture vessels are of two-phase blood culture systems, the so-called Gibco-Bio-Cult systems with several nutrient media known, which have a special safety seal and a partition for separating liquid and solid nutrient media. These blood culture vessels avoid the inoculation of samples for subcultures at different time intervals and thus the repeated opening of the vessels. The blood-mixed liquid nutrient medium flows through rotation of the vessel into the intended chamber, flushes and inoculates the solid agar phase arranged on the partition wall and is then poured back into the intended chamber in the vessel.

The disadvantages of this two-phase blood culture system are that no direct connection to the venous blood collection system is possible due to the construction of the vessel with a relatively large volume. The blood draw from the vein and consequently the blood supply to these blood culture vessels or vials takes place mainly by means of syringes, only for small vessel volumes by means of hose connections for connection to the drain cannula or a blood draw cutlery. Although these two-phase blood culture vials allow a 448,737 faster growth of the colonies on agar, there is an increased risk of contamination due to the separate blood draw. Furthermore, even in this blood culture system, the optical production of colony formations during the incubation and consequently, especially in infant cultures, the microbiological examination is more difficult. Therefore, in certain blood culture species, it is necessary to rinse the solid agar phase at specified intervals repeatedly with liquid nutrient medium-blood mixture.

According to DE-OS 2221096 and DE-OS 2332133, blood collection devices are known as blood collection instruments and as cannula units for taking blood samples.

According to the said blood collection methods, the blood collection instrument consists of a cannula with holder, the inside end section of the cannula being surrounded by a valve with a ventilation device. The cannula valve can be brought from a position in which it allows aeration through a narrow opening to enable blood indication to a further position in which it prevents the passage of air or blood through the opening.

The holder consists of a tubular main part and a part with an inner conical surface. The tubular part serves to receive the evacuated blood culture vessel with a pierceable closure. The conical surface serves as a stop for the culture vessel, when the closure piercing the end of the cannula has pierced it.

Blood collection and blood culture systems are connected through these holders in a known manner.

Similarly, the cannula unit protected according to DE-OS 2332133 is constructed, in which the front part of the cannula is inserted into the vein and the underlying cannula part in a blood culture collection container. This cannula unit is characterized by a one-way valve to prevent the backflow of fluid from the collection container into the patient's bloodstream and by means of visible blood flow indication.

The cannula also has a holder adapted to the shape of the blood culture collection vessel, which for receiving this vessel is preferably designed as a perforated cylinder.

After venipuncture, blood collection and blood culture systems are also connected via the holder. Like the above-mentioned, this loose connection also has the disadvantages of increased contamination of the entire system as well as cumbersome and expensive equipment technical handling.

From DE patent ll0357 vacuum ampoules for sterile draining of blood and other liquids are known, in which the ampoule neck closed at the top is pulled over the stopper and cannula supported by the cannula and pressed from the lower end of the ampoule and which is melted under high vacuum at the lower end.

These vacuum ampoules are used exclusively for blood collection; the aspirated blood must then be introduced into a separate blood culture system. To replenish this blood donation system, nutrient media are required, which during cultivation offer the bacteria favorable conditions for their development. The corpuscular and non-corpuscular defense forces present in the blood are capable of killing resp. inactivate large amounts of bacteria. For carrying out blood tests by means of cultures of bacteria from the blood, a variety of substances and substances are known, which inhibit these defense forces. 448 737 Thus, a polyanethol sulfonate is known which has been found to be best suited for these purposes. It also exhibits anticoagulant properties. However, the known media are not suitable for two-phase blood collection systems, as they have too few antibacterial properties and are not able to make the bacterial colonies present in the blood collection system after culture visible. The object of the invention is to provide a method and a device for examining bacterial mites, in which the disadvantages of the known methods and devices are eliminated, the safety before contamination is significantly increased, the blood collection and the establishment of bacterial cultures is simplified and the hitherto high appa. - steering technology consumption of blood collection and blood culture systems is avoided.

The object of the invention is therefore to develop a method for examining bacterial chemicals, according to which the time and path from the discharge of blood from the vein to its effect on liquid and solid blood nutrient cultures is considerably shortened, and to provide a device, in which all the necessary functions of the blood collection and culture system are combined into as small a number of components as possible with an appropriate structure, and to design the nutrient medium so that in the collection and examination system anticoagulant properties and antibacterial activity against blood properties are developed.

According to the invention, the method for detecting bacterial ants, in which the blood is via a blood collection system in large doses, is sucked into a vessel provided with a vacuum 448 737 and / or a special gas filling in a blood culture system and thereby mixed with a liquid nutrient medium, in that the vessel sucked in the blood simultaneously inoculates the liquid and thus mixes the solid nutrient medium. At a certain temperature and inclination of the vessel in the blood culture system, the bacterial culture is then generated in both nutrient media and made visible by an indicator dye included in the nutrient media.

The device according to the invention for examining bacterial mites, consisting of a blood collection and a blood culture system, is characterized in that the vessel in the blood culture system is designed as an ampoule with an annular contraction and that this annular contraction position on the length of the ampoule is adapted to the required liquid nutrient medium, preferably the ratio 1: 3.

Furthermore, the arrangement is characterized in that the liquid and the solid nutrient medium are arranged one after the other in the vessels of the blood culture system and that they are in contact with each other at a position of the vessel in the angular range 0 - 1350 and that the volume and position of the solid nutrient medium are fixed by the annular the contraction.

The annular contraction has a permanent break on its surface, which enables an even, rupture and shatterproof separation of the two parts of the blood culture vessel. The diameter of the passage opening of the annular contraction is then preferably in the range 6 <.10 mm, since the amount and position of the solid agar phase is determined by the starting point for transverse fl f, the radius of the section contraction.

The device according to the invention is further characterized by 448 757 that the vessels of the blood culture system are connected to the blood collection system via an elastic connecting piece, preferably of a transparent soft plastic material, which is a closed unit located one behind the other and that the axis of the blood culture system is arranged at an angle to the blood culture system. and moving in any direction.

The blood collection system according to the invention is characterized in that the lower part of the tube of the injection cannula is so elongated that the tube encloses the interruptible glass tip of the vessels of the blood culture system, whereby the interruption of the glass tip is made possible before the blood draw.

In addition, around the blood culture system's interruptible tip, the elastic connector and the blood collection system with a fitting, a cover is arranged to protect the blood collection system.

The nutrient medium used in the method according to the invention for the detection of bacterial ants consists of a liquid and a solid part containing antibacterial substances, substances for preventing blood coagulation and a pH-dependent indicator system; preferably the liquid part of the nutrient medium contains the substances indomethacin and carrageenan.

Exemplary embodiments The advantages of the method and the device according to the invention consist in that a combination of several hitherto different procedural steps is achieved by simultaneous qualitative increase of the blood collection and examination method by means of a contamination-safe, easy-to-handle device. . 448 757 10 In addition, by using high-quality nutrient media with antibacterial additives and solid nutrient media with indicator dyes, it is possible to extract a smaller blood volume and thereby establish bacterial cultures.

Depending on the composition of the nutrient medium with these particular additives, it is possible to grow only microbes, i.e. to extract a small amount of blood and thereby keep the two parts of the vessel in the blood culture system to volume optimally small compared to known blood culture vessels. Through the simultaneous inoculation of the liquid and solid nutrient medium and through the optical production of the bacterial growth, a significant simplification of the entire procedure and a reduction of the bacteriological workload takes place.

The direct blood supply in the two-phase blood culture vessel, the rapid inoculation of the two nutrient media after the blood draw and the incubation of bacterial cultures in the two-part blood culture vessel without further filling and thus without the possibility of contamination also guarantees a rapid and safe diagnosis.

The combination of liquid and solid nutrient medium in the blood culture vessel according to the invention makes it possible for stimulating the bacterial growth of certain colonies to more or less moisten the surface of the solid agar phase at certain time intervals through the liquid level in the liquid nutrient medium at oblique position of the vessel.

By the method according to the invention, the separate subcultivation from the blood culture vessel and thus its procedural and apparatus technical costs can be eliminated. The annular contraction, after bacterial growth has occurred, allows an easy and safe separation of the parts of the blood culture vessel for further examination of the two nutrient media isolated from each other. The tension ring of the annular contraction enables an even interruption at reduced risk of injury.

The method and the device according to the invention will in the following be illustrated and explained on the basis of an exemplary embodiment. An example is shown in the accompanying drawing, in which Figures 1 and 2 show a longitudinal section of the embodiment according to the invention in schematic representation.

In the procedure for examining bacteria in the blood, a blood sample is aspirated from a patient's vein via a blood collection system, a cannula or syringe system of a specific construction, mass-metered into a glass culture vessel of vacuum or a special gas filling and then with the liquid nutrient medium contained in the vessel. The absorbed blood then inoculates the liquid nutrient medium provided with antibacterial additives, whereby at the same time a solid nutrient medium is inoculated into the blood culture vessel through the V nutrient medium-blood mixture. Thereafter, the bacterial culture is generated in both nutrient media at a certain temperature and inclination of the vessel - depending on the bacterial culture intended for examination. By adding indicator indicator dye, which is preferably added to the solid nutrient medium, it is possible to indicate increasing bacteria in both media.

The device according to the invention consists of a blood donor- .._......_._...._...._.- _ ”. __ 448 757 12 and a blood culture system. The blood culture system consists of a two-part vitreous, the blood culture vessel 1; 2, which according to the invention is designed as an ampoule - consisting of part 1 and part 2. This vessel 1; 2 has an annular contraction 3, which divides its length and volume in a certain ratio of 1: 3. W On the surface of the annular contraction 3 a permanent tension ring 9 is arranged on its circumference, which by a defined tension in the glass allows a clean and simple division of the blood culture vessel 1; 2 both parts with less risk of injury. The diameter of the annular contraction 3 depends on the diameters of the passage opening for the debug loop and is in the range 6 <10 mm.

The liquid and solid nutrient medium are in the blood culture vessel 1; 2 arranged one after the other and are in contact with each other, by in part 2 of the blood culture vessel 1; 2 a solid agar 10 is included and in the part 1 a liquid nutrient medium 11. The annular contraction 3 fixes the volume and position of the solid agar by holding it by the curvature of the contraction.

The volume and thus the size of the blood culture vessel 1; 2 are variable in relation to the amount of blood to be removed. Through the composition of nutritional; the media with certain antibacterial additives against the body's own defenses, according to the invention 17 'it is possible to take a small amount of blood and thereby maintain a small volume and dimension of the vessel 1; 2 since already a small amount of microbes for cultivation in blood- V! the cultures show visible reactions. The blood culture vessel 1; 2 is preferably designed as an ampoule with a detachable glass tip 4, which is enclosed by an elastic connecting piece 5, e.g. a connecting hose of transparent soft plastic such as PVC hose type 60 S ready, so that thereby the blood culture system is connected to the blood collection system as a closed unit located in succession.

Through the elastic connecting piece 5, the axis of the blood collection system is arranged at an angle to the axis of the blood culture system and movable in any direction.

The blood collection system consists of the injection cannula 6, which is enclosed by a glass jacket 7 and in the lower part extended so that this tubular extension encloses the blood culture vessel 1; 2 breakable glass tip 4.

Around the blood culture vessel l; 2 can be broken off, the elastic connecting piece 5 and the blood collection system are fitted with a cover 8.

Room 12 is evacuated or for the cultivation of special bacterial cultures provided with a gas atmosphere with defined pressure. To puncture the vein, the cover 8 is removed before the use of the device according to the invention and the glass jacket 7 I of the injection cannula 6 is sawn and interrupted, so that the sterile cannula 6 is exposed. After the insertion into the vein, a cutting movement of the blood culture system around the axis of the blood collection system is performed at the lying cannula 6, until the glass tip 4 is broken off. By releasing the action of the vacuum, a negative pressure is generated in the cannula 6, so that according to the present vacuum a certain blood sample in a metered dose is automatically sucked into the vessel 1 of the blood culture system; 2.! Simultaneously with this mass-dosed aspiration of blood, the inoculation of the liquid nutrient medium II and thereby mixed with the solid agar phase 10. 448 737 14 After removal of the injection cannula 6 from the vein and its closure with a sterile plug, preferably of plastic, is carried out at a determined oblique position of the whole blood culture system the incubation, i.e. the cultivation of the local media 10 and ll at temperatures preferably in the range 35 ... 370.

A certain inclination of the vessel 1; 2 of the blood culture system is, depending on the volume of the vessel and thus the solid and liquid nutrient medium 10; 11, necessary to cause the liquid level 11 of the liquid nutrient medium to partially cover the surface of the solid agar 10.

By inoculating the nutrient media 10; ll there is an increase in the bacteria; According to the invention, the growth of bacterial cultures in both media becomes visible through the color change of an indicator dye, in addition to these. The microbiological examination is carried out separately, by scratching the blood collection and culture system and breaking it apart by breaking off parts 1 and 2 in the vessel 1 of the blood culture system; 2 at the refraction 9 of the annular contraction 3. Thereby a division of the liquid medium 11 takes place, from the bacterial-grown solid medium 10, so that a separate inoculation and sampling is possible for further bacteriological processing resp. examination of both media.

The separation of the liquid nutrient medium 11 from the solid agar phase 10 is effected upon interruption of the vessel 1; 2 in the blood culture system in such a way that part 2 with agar 10 points upwards. Thereby it is also possible 9 / that to the vessel 1; 2 set certain gas fillings, with which bacterial strains can increase, which in oxygen atmosphere I * can only be detected with difficulty. 448 737 The liquid and the solid part of the nutrient medium have the following preferred composition for the purpose of the invention: Liquid part of the nutrient medium: Distilled water Sucrose Pancreatic peptone Glucose Meat extract Sodium chloride Indomethacin 0.0028 Carrageenan 0.01 ...

Solid portion of the nutrient medium: Sterile nutrient agar Glycos 1000 100 20 10 2.5 3.0 g ... 0.0035 g 0.08 g LQKIUIIIQKI I '- DAB7 99.5 g 015 g Bromocresol purple (indicator) 0.0076 g De individual constituents.channel depending on the purpose of use also be present in other proportions.

Claims (1)

A patent for detecting bacterial mites, in which the blood is sucked via a blood collection system in a metered dose into a vessel equipped with a vacuum and / or a special gas filling in a blood culture system and thereby mixed with a liquid nutrient medium, characterized in that it the blood sucked into the vessel simultaneously inoculates the liquid nutrient medium provided with antibacterial additives and mixed with a solid nutrient medium and then at a certain temperature and inclination of the vessel in both nutrient media the bacterial culture is generated and made visible by the addition of indicator dye. Device for examining bacterial ants consisting of a blood collection and a blood culture system, characterized in that the vessel in the blood culture system (1; 2) is designed as an ampoule with an annular contraction (3) and that the annular contraction ( 3) position on the length of the ampoule is adapted to the required amount ratio between a solid and a liquid nutrient medium (10; 11), preferably in the ratio 1: 3. Device according to claim 2, characterized in that the annular contraction (3) has on its surface a break ring (9) and the diameter of its passage opening is in the range 6 <10 m. Device according to claim 2, characterized in that it liquid and the solid nutrient medium (10; 11) are arranged one after the other in the vessels of the blood culture system (1; 2) and are in contact with each other and the solid nutrient medium (10) is fixed in position by the annular contraction (3). u! s; Device according to claim 2, characterized in that the vessels of the blood culture system (1; 2) are connected to the blood collection system via an elastic connecting piece (5), preferably of a transparent soft plastic material as a closed unit located one after the other and that the axis of the blood collection system is arranged at an angle to the axis of the blood culture system and movable in each direction. Device according to claim 2, characterized in that the lower part of the tube of the injection cannula (6) is so elongated that it encloses the interruptible glass tip (4) of vessels of the blood culture system (1; 2). Device according to claims 2 - 6, characterized in that a cover (8) is arranged around the breakable glass tip (4) of the blood culture system, the elastic connecting piece (5) and the blood collection system with a fitting.