CN1635109A - Anaerobic bacteria culturing installation and method - Google Patents
Anaerobic bacteria culturing installation and method Download PDFInfo
- Publication number
- CN1635109A CN1635109A CN 200410090290 CN200410090290A CN1635109A CN 1635109 A CN1635109 A CN 1635109A CN 200410090290 CN200410090290 CN 200410090290 CN 200410090290 A CN200410090290 A CN 200410090290A CN 1635109 A CN1635109 A CN 1635109A
- Authority
- CN
- China
- Prior art keywords
- vessel
- sample
- bacteria
- cultivated
- interval
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an apparatus special for culturing anaerobe and a method therefore. The apparatus is composed of vessels inosculating with each other, and each vessel is equipped with an isolation fastener on the inside circumference, a ring seal pad is arranged in the space between the fastener and corresponding vessel outer wall. The method includes the following steps: (1) preparing a substance (a specimen) for culture; (2) culturing in the anaerobic environment in the anaerobe culture apparatus; (3) culturing the specimen of the step 1 in the normal aerobic environment; (4) judging. The anaerobe culture apparatus has advantages of simple structure, simple operation, practicality, it can not only provide direct, effective, economical diagnose standards for the patients, but also provide precise, reliable anaerobe culture results. The anaerobe cultured by the method has high positive rates and reliable results, the method is suitable for the anaerobe culture in big, medium, small microbe laboratories, and costs low.
Description
Technical field:
The present invention relates to the device and method of a kind of special cultivation anerobe, especially relate to the device and method that the severe oxygen bacterium of a kind of usefulness carries out anaerobic bacteria culture.
Background technology:
Much more comparatively anerobe infects the body case clinically and to see, relevant anaerobic bacteria culture is in field of medical examination, people are puzzling always, especially poor big of medical condition, in, there is bigger difficulty in small hospital to solving anaerobic bacteria culture, most clinically at present anaerobism bags with ready-made purchase, anaerobic jar, the anaerobic box culture method, it all is to cause the anaerobic environment through method physics or chemistry that various anaerobism are cultivated, and the employing anaerobic jar, its working routine of anaerobism bag cultivation anerobe is loaded down with trivial details, manufacture complexity, the requirement condition harshness, it is low that part is cultivated positive rate, common laboratory particularly middle-size and small-size to some and basic hospital can not independently be finished the cultivation of anerobe, and being difficult to reach needs standard.Develop into present state-of-the-art anaerobic box and cultivate anerobe by traditional anaerobic jar, anaerobism bag cultivation anerobe now, but because of the cost height of anaerobic box own, expense costliness, because of regional disparity, patient's ability to shoulder economically difference, so advanced medical facilities are difficult to promote the use of at large.
Summary of the invention:
The object of the present invention is to provide a kind of simple in structure, working method is easy, economical and practical, suitable extensively promote, can not only give clinically provide direct, effective, economic diagnosis index, and can improve the device of novel anaerobic bacteria culture of the positive rate of anerobe.
Another object of the present invention is to provide a kind of anerobe positive rate height of turning out, reliable results, method is easy, and the anaerobic bacteria culture, the expense that are fit to large, medium and small Microbiological Lab are low, simple and convenient and easy to study, is convenient to the method for the novel anaerobic bacteria culture of penetration and promotion.
First purpose of the present invention is implemented by following technical scheme: it is by last, the vessel that coincide mutually constitute down, periphery in arbitrary vessel of described upper pot or lower pot is provided with the isolation fastener that forms a circle, and the height of described isolation fastener is lower than the height of place vessel, form an interval between the outer wall of described isolation fastener and corresponding vessel, bottom in described interval is provided with Gask-O-Seal, the top edge of described another vessel is buckled on the Gask-O-Seal of the described vessel that are provided with Gask-O-Seal, described on, lower pot adopts screw thread or clamping to connect.
Described upper and lower part vessel be shaped as circle, ellipse, tetragon, Polygons or abnormity.
Described isolation fastener be shaped as circle, ellipse, tetragon, Polygons or abnormity.
The edge, outer ring of described Gask-O-Seal is consistent with the bottom shape of the described vessel that are provided with Gask-O-Seal, and the inner ring edge of described Gask-O-Seal is consistent with the shape of described isolation fastener.
The height of described upper and lower part vessel is 5-25mm, and the floorage of described upper and lower part vessel is 15cm
2-140cm
2, and be lower than the height of described another vessel at the height that is provided with the vessel of isolating fastener and Gask-O-Seal.
Be provided with the division board that encloses into a circle in vessel in described upper pot or lower pot or two vessel, described division board is divided into two intervals with the place vessel---interior interval and outer interval, the interval is the interval in the described division board circle in described, described outer interval is the outer interval of described division board circle, and the height of described division board will be lower than the height of its place vessel.
Be provided with dividing plate in any vessel of described upper pot or lower pot, described dividing plate is divided into isolated two intervals mutually with its described vessel, and described dividing plate is lower than the height of place vessel.
Another object of the present invention is implemented by following technical scheme: include following steps:
(1), at first prepares material---sample to be cultivated;
(2), step (1) gained sample being carried out oxygen-free environment in the device of described novel anaerobic bacteria culture cultivates;
The solid medium that has soon prepared places the vessel of the device of described novel anaerobic bacteria culture, cool off, solidify, the known severe bacteria of inoculation in filling vessel of described solid medium, inoculation sample to be cultivated in filling another vessel of described solid medium;
Or put into the solid medium that has prepared in the interior interval of the described upper and lower vessel in the device of described novel anaerobic bacteria culture, cool off, solidify, the known severe bacteria of inoculation on the interval solid medium in vessel is inoculated sample to be cultivated on the interval solid medium in described another vessel;
Or in the interior interval of arbitrary vessel of described upper pot in the device of described novel anaerobic bacteria culture or lower pot and outside the interval put into the solid medium that has prepared, cool off, solidify, the known severe bacteria of inoculation on the solid medium in interval in described, inoculation sample to be cultivated on the solid medium in interval outside described;
Or arbitrary vessel of the described upper and lower vessel in the device of described novel anaerobic bacteria culture, by putting into the solid medium that has prepared respectively in mutual isolated two intervals that dividing plate separated, cool off, solidify, the known severe bacteria of inoculation on the interval solid medium in the described part that separates by described dividing plate, inoculation sample to be cultivated on the interval solid medium in the described another part that separates by described dividing plate;
Described upper pot and described lower pot screwing hermetic are isolated with extraneous air, and the spacing between the described solid medium in wherein said upper pot and the described lower pot is at least greater than 5mm; Then the device of described novel anaerobic bacteria culture is put into 37 degrees centigrade of constant incubators and cultivated 10-18h, observe the growth of the bacterium of described inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
(4), by above three bacterial colonys that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultative any type of detesting bacteria, severe bacteria or not having bacterial growth.
Step (4) is to judge by the following method:
(A), sample described to be cultivated carried out oxygen-free environment cultivate in the device of novel anaerobic bacteria culture, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is severe bacteria, or does not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may be for detesting bacteria or facultatively detesting bacteria;
(B), sample described to be cultivated is cultivated on the common substratum that has under the foster environment, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is for detesting bacteria, or do not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may or facultatively be detested bacteria for severe bacteria;
Information summary by above-mentioned steps (A), (B) bacterial growth obtains four kinds of detected results,
(I), by (A, a) and (B, a) to obtain the result be no bacterial growth;
(II), by (A, a) and (B, b) to obtain the result be severe bacteria, promptly this sample is that severe bacteria infects;
(III), by (A, b) with (B, a) obtain the result and detest bacteria, promptly this sample infects for detesting bacteria;
(IV), to obtain the result by (A, b) and (B, b) be facultative severe bacteria, promptly this sample is that facultative severe bacteria infects.
Sample described to be cultivated is human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.
Described solid medium is that solid-based basal culture medium, solid nutrient medium, solid are selected any of substratum, solid differential medium.
This method can be used as the drug sensitivity test of detesting bacteria.
The described thickness of putting into the off-the-shelf solid medium of upper and lower part vessel is at least 3mm.
Advantage of the present invention is: the apparatus structure of novel anaerobic bacteria culture is simple, working method is easy, and is economical and practical, and clinical direct, effective, the economic Case definition that provides can not only be provided in suitable extensively popularization, accurate, reliable to the cultivation results of anerobe simultaneously.
The anerobe positive rate height that the method for novel anaerobic bacteria culture is turned out, reliable results, method is easy, and the anaerobic bacteria culture, the expense that are fit to large, medium and small Microbiological Lab are low, simple and convenient and easy to study, are convenient to penetration and promotion.Method with biological oxygen consumption is created oxygen-free environment, replaces the method for physics, chemistry, and very big effect is played in the cultivation that popularizes anerobe.Utilize severe oxygen bacterium in the growth and breeding process with microenvironment in oxygen depletion produce a large amount of CO
2, cause anaerobic environment, help the growth and breeding of anerobe.Along with microbiological development, serial anaerobic bacteria culture relevant item is brought in constant renewal in, and the economical load of having avoided existing cultural method to cause to patient priorly causes difficulty and loss for clinical diagnosis, treatment.And novel method is simple, and is economical and practical, is suitable for hospitals at different levels, R﹠D institution, and experiment is accurate, reliable, suitability is strong.
The sample for the treatment of cultivation by this method can be by cultivating anerobe, thereby the sample that can treat cultivation is more accurately identified, its reliable results, and method is easy, the kind that is fit to large, medium and small Microbiological Lab is identified, expense is low, simple and convenient and easy to study, is convenient to penetration and promotion.This method can be used as the drug sensitivity test of detesting bacteria simultaneously.
The drawing explanation of accompanying drawing:
Fig. 1 is the one-piece construction sectional view of the device embodiment 1 of the novel anaerobic bacteria culture of the present invention.
Fig. 2 is the vertical view of the upper pot of Fig. 1.
Fig. 3 is the vertical view of the lower pot of Fig. 1.
Fig. 4 is the one-piece construction sectional view of the device embodiment 2 of the novel anaerobic bacteria culture of the present invention.
Fig. 5 is the vertical view of the upper pot of Fig. 4.
Fig. 6 is the vertical view of the lower pot of Fig. 4.
Fig. 7 is the one-piece construction sectional view of the device embodiment 3 of the novel anaerobic bacteria culture of the present invention.
Fig. 8 is the vertical view of the upper pot of Fig. 7.
Fig. 9 is the vertical view of the lower pot of Fig. 7.
Figure 10 is the orthohexagonal one-piece construction sectional view that is shaped as of the device embodiment 4 upper and lower part vessel of the novel anaerobic bacteria culture of the present invention.
Figure 11 is the vertical view of the upper pot of Figure 10.
Figure 12 is the vertical view of the lower pot of Figure 10.
Figure 13 is the one-piece construction sectional view that is shaped as circle of the device embodiment 4 upper and lower part vessel of the novel anaerobic bacteria culture of the present invention.
Figure 14 is the vertical view of the upper pot of Figure 13.
Figure 15 is the vertical view of the lower pot of Figure 13.
Figure 16 treats the general flow chart of decision method of the material bacterium kind of cultivation for the present invention.
Embodiment:
Embodiment 1: as Fig. 1, Fig. 2, shown in Figure 3, the device of novel anaerobic bacteria culture is made of the upper and lower vessel that coincide mutually, establish the isolation fastener 3 that forms a circle at the periphery of upper pot 1, the height of isolating fastener 3 is lower than the height of upper pot 1, form an interval between the outer wall of isolation fastener 3 and corresponding upper pot, the bottom in interval is provided with Gask-O-Seal 4.The top edge 9 of lower pot 2 is buckled on the Gask-O-Seal 4 of upper pot 1, and upper and lower part vessel 1,2 adopt screw thread or clamping to connect.The bottom shape of upper pot 1 and lower pot 2 is circular.The shape of isolating fastener 3 and Gask-O-Seal 4 also is circular.Upper pot 1 adopts screw thread or clamping to be connected with lower pot 2.The height of upper pot 1 is 5mm, and the height of lower pot 2 is 15mm.The internal diameter of upper pot is 110mm, and the lower pot external diameter is 105mm.
Can in upper pot 1 and lower pot 2, put into off-the-shelf solid nutrient medium respectively in use, cool off, solidify, inoculate sample to be cultivated on the solid medium of vessel therein then, the known severe bacteria of inoculation on the solid medium of another vessel, then that upper and lower vessel lid is tight, sealing and extraneous air have been convenient to the cultivation of sample.Wherein the spacing between the solid medium in upper pot and the lower pot is at least greater than 5mm.
Embodiment 2: as Fig. 4, Fig. 5, shown in Figure 6, the device of novel anaerobic bacteria culture is made of the upper and lower vessel that coincide mutually, establish the isolation fastener 3 that forms a circle at the periphery of lower pot 2, the height of isolating fastener 3 is lower than the height of lower pot 2, form an interval between the outer wall of isolation fastener 3 and corresponding lower pot 2, the bottom in interval is provided with Gask-O-Seal 4.The top edge 10 of upper pot 1 is buckled on the Gask-O-Seal 4 of lower pot 2, and upper and lower part vessel 1,2 adopt clamping to connect.Upper pot 1 and lower pot 2 be shaped as ellipse.Isolate the rectangle that is shaped as of fastener 3, the outer ring edge shape of Gask-O-Seal 4 is consistent with the shape of lower pot 2, promptly also is oval, and the inner ring edge shape of Gask-O-Seal 4 is consistent with the shape of isolating fastener 3, is rectangle.And Gask-O-Seal 4 closely contacts with isolation fastener 3 with the periphery of lower pot 2, and upper pot 1 adopts clampings to be connected with lower pot 2.The height of upper pot 1 is 25mm, and upper pot 1 is 55mm in long axis length, and minor axis length is 45mm, and the height of lower pot 2 is 15mm, and lower pot 2 is 60mm in long axis length, and minor axis length is 50mm.In two vessel in upper pot 1 and the lower pot 2, be provided with the division board 5 that encloses into a circle, division board 5 is divided into two intervals with the place vessel---and interior interval 6 and outer interval 7, interior interval 6 is the intervals in division board 5 circles, outer interval 7 is the outer intervals of division board 5 circles, and the height of division board 5 will be lower than the height of its place vessel.
In use can upper pot 1 and lower pot 2 interior interval 6 in put into off-the-shelf solid medium respectively, cool off, solidify, inoculate sample to be cultivated on interior interval 6 the solid medium of vessel therein then, the known severe bacteria of inoculation on interval 6 the solid medium in another vessel, then that upper and lower vessel lid is tight, sealing and extraneous air have been convenient to the cultivation of sample.Wherein the spacing between the solid medium in upper pot and the lower pot is 8mm.
Embodiment 3: as Fig. 7, Fig. 8, shown in Figure 9, the device of novel anaerobic bacteria culture is made of the upper and lower vessel that coincide mutually, establish the isolation fastener 3 that forms a circle at the periphery of upper pot 1, the height of isolating fastener 3 is lower than the height of upper pot 1, form an interval between the outer wall of isolation fastener 3 and corresponding upper pot 1, the bottom in interval is provided with Gask-O-Seal 4.The top edge 9 of lower pot 2 is buckled on the Gask-O-Seal 4 of upper pot 1, and upper and lower part vessel 1,2 adopt clamping to connect.The bottom shape of upper pot 1 and lower pot 2 is a square.The shape of isolating fastener 3 and Gask-O-Seal 4 also is a square.Upper pot 1 adopts clamping to be connected with lower pot 2.The height of upper pot 1 is 12mm, and the upper pot 1 bottom surface length of side is 80mm, and the height of lower pot 2 is 22mm, and the lower pot 1 bottom surface length of side is 75mm.In upper pot 1, be provided with the division board 5 that encloses into a circle, division board 5 is divided into two intervals with the place upper pot---and interior interval 6 and outer interval 7, interior interval 6 is the intervals in division board 5 circles, outer interval 7 is the outer intervals of division board 5 circles, and the height of division board 5 will be lower than the height of its place upper pot 1.
Can in interior interval 6 and outer interval 7 of upper pot 1, put into off-the-shelf solid medium respectively in use, cool off, solidify, on interior interval 6 solid nutrient medium, inoculate sample to be cultivated then, inoculate known severe bacteria on interval 7 the solid nutrient medium outside, then that upper and lower vessel lid is tight, sealing and extraneous air have been convenient to the cultivation of sample.Wherein the spacing between the solid medium in upper pot and the lower pot is 12mm.
Embodiment 4: as Figure 13, Figure 14, shown in Figure 15, the device of novel anaerobic bacteria culture is made of the upper and lower vessel 1,2 that coincide mutually, establish the isolation fastener 3 that forms a circle at the periphery of lower pot 2, the height of isolating fastener 3 is lower than the height of lower pot 1, form an interval between the outer wall of isolation fastener 3 and corresponding lower pot 2, the bottom in interval is provided with Gask-O-Seal 4.The top edge 10 of upper pot 1 is buckled on the Gask-O-Seal 4 of lower pot 2, and upper and lower part vessel 1,2 adopt clamping to connect.The shape of upper pot 1 and lower pot 2 can be circle, and as Figure 10, Figure 11, shown in Figure 12, the shape of upper pot 1 and lower pot 2 can be regular hexagon, and the length of side is 35mm.The shape of upper pot 1 and lower pot 2 also can be abnormity, as heart, cartoon shape etc.The shape of isolating fastener 3 and Gask-O-Seal 4 is circle, and the height of lower pot 2 is 10mm, and the internal diameter of lower pot 2 bottom surfaces is 90mm, and the height of upper pot 1 is 20mm, and the external diameter of upper pot 1 bottom surface is 86mm.Be provided with dividing plate 8 in upper pot 1, the height sum of the height of dividing plate 8 and isolation fastener 3 is less than the height of place lower pot 2 at least.
During use, in mutual isolated two portions that upper pot 1 is separated by dividing plate 8, put into off-the-shelf solid medium respectively, cool off, solidify, the known severe bacteria of inoculation on the solid nutrient medium in the part of the vessel that separate by dividing plate 8, inoculation sample to be cultivated on the solid nutrient medium of the vessel of the another part that separates by dividing plate; Then that upper and lower vessel lid is tight, sealing and extraneous air have been convenient to the cultivation of sample.
Embodiment 5: the method for novel anaerobic bacteria culture includes following steps:
(1), at first prepare material---sample to be cultivated, as human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.Sample can not have pollution.The sample that takes off will advance fast cultivation, sample is not placed for a long time, prevents to detest dead and other pollutions of bacteria.There is not bacterial growth in the foster substratum of detesting of usefulness before cultivating.
(2), step (1) gained sample being carried out oxygen-free environment in the device of embodiment 1 cultivates; Be about to the upper and lower part vessel that solid nutrient medium places the device of embodiment 1, the known severe bacteria of inoculation on the solid nutrient medium of vessel, inoculation sample to be cultivated on the solid nutrient medium of another vessel, with upper pot and lower pot screwing hermetic, isolate with extraneous air, wherein between the nutritional medium in upper pot and the lower pot spacing is arranged, its spacing is at least greater than 5mm; Then novel anaerobic bacteria culture device is put into 37 degrees centigrade of constant incubators and cultivate 12h, observe the growth of the bacterium of described inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
By above three bacterial colonys that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultative any type of detesting in bacteria, severe bacteria or the non-bacterial infection.
Treat the decision method of the material bacterium kind of cultivation:
(A), sample that will be to be cultivated carries out oxygen-free environment and cultivates in the device that carries out anaerobic bacteria culture with severe oxygen bacterium, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is severe bacteria, or does not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may be for detesting bacteria or facultatively detesting bacteria;
(B), same sample to be cultivated is cultivated on the common substratum that has under the foster environment, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is for detesting bacteria, or do not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may or facultatively be detested bacteria for severe bacteria;
Information summary by above-mentioned steps (A), (B) colony growth obtains four kinds of detected results,
(I), by (A, a) and (A, a) to obtain the result be no bacterial growth, and promptly this sample is a non-bacterial infection;
(II), by (A, a) and (B, b) to obtain the result be severe bacteria, promptly this sample is that severe bacteria infects;
(III), by (A, b) with (B, a) obtain the result and detest bacteria, promptly this sample infects for detesting bacteria;
(IV), to obtain the result by (A, b) and (B, b) be facultative severe bacteria, promptly this sample is that facultative severe bacteria infects.
Then the bacterial classification that obtains is further carried out biochemical identification.This method can be used as the drug sensitivity test of detesting bacteria simultaneously.
Embodiment 6: the method for novel anaerobic bacteria culture includes following steps:
(1), at first prepare material---sample to be cultivated, as human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.Sample can not have pollution.The sample that takes off will advance fast cultivation, sample is not placed for a long time, prevents to detest dead and other pollutions of bacteria.No bacterial growth in the substratum is supported in standby detesting before cultivating.
(2), step (1) gained sample being carried out oxygen-free environment in the device of embodiment 2 cultivates; Promptly put into the solid differential medium in the interior interval of the upper and lower vessel in the device of novel anaerobic bacteria culture, the known severe bacteria of inoculation on the solid differential medium in interval in vessel, inoculation sample to be cultivated on the solid differential medium in interval in another vessel, with upper pot and lower pot screwing hermetic, isolate with extraneous air, wherein the spacing between the solid differential medium in upper pot and the lower pot is 8mm; The device that severe oxygen bacterium is carried out anaerobic bacteria culture is put into 37 degrees centigrade of constant incubators and is cultivated 10h then, observes the growth of the bacterium of inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
By above three bacterium colonies that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultative any type of detesting in bacteria, severe bacteria or the non-bacterial infection.
Method such as embodiment 5 that its sample bacterium kind for the treatment of cultivation is identified.Then the bacterial classification that obtains is further carried out biochemical identification.
This method can be used as the drug sensitivity test of detesting bacteria.
Embodiment 7: the method for novel anaerobic bacteria culture includes following steps:
(1), at first prepare material---sample to be cultivated, as human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.Sample can not have pollution.The sample that takes off will advance fast cultivation, sample is not placed for a long time, prevents to detest dead and other pollutions of bacteria.There is not bacterial growth in the foster substratum of detesting of usefulness before cultivating.
(2), step (1) gained sample being carried out oxygen-free environment in the device of embodiment 3 cultivates; In use can upper pot 1 interior interval 6 and outer interval 7 in put into solid selection substratum respectively, select inoculation sample to be cultivated on the substratum at interior interval 6 solid then, interval 7 interior solids are selected the known severe bacteria of inoculation on the substratum outside, with upper pot and lower pot screwing hermetic, isolate with extraneous air, wherein the spacing between the nutritional medium in upper pot and the lower pot is at least greater than 10mm; The device that severe oxygen bacterium is carried out anaerobic bacteria culture is put into 37 degrees centigrade of constant incubators and is cultivated 18h then, observes the growth of the bacterium colony of inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
By above three bacterium colonies that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultatively detest in bacteria, severe bacteria or the non-bacterial infection any.
Method such as embodiment 5 that its sample bacterium kind for the treatment of cultivation is identified.Then the bacterial classification that obtains is further carried out biochemical identification.
Embodiment 8: the method for novel anaerobic bacteria culture includes following steps:
(1), at first prepare material---sample to be cultivated, as human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.Sample can not have pollution.The sample that takes off will advance fast cultivation, sample is not placed for a long time, prevents to detest dead and other pollutions of bacteria.There is not bacterial growth in the foster substratum of detesting of usefulness before cultivating.
(2), step (1) gained sample being carried out oxygen-free environment in the device of embodiment 4 cultivates; Promptly in the device of novel anaerobic bacteria culture, in the lower pot that dividing plate separated, put into the solid-based basal culture medium respectively in mutual isolated two portions, the known severe bacteria of inoculation on the solid-based basal culture medium of a part of vessel that separate by dividing plate, inoculation sample to be cultivated on the solid-based basal culture medium of another part vessel that separate by dividing plate; With upper pot and lower pot screwing hermetic, isolate with extraneous air, the device that severe oxygen bacterium is carried out anaerobic bacteria culture is put into 37 degrees centigrade of constant incubators and is cultivated 15h then, observes the growth of the bacterium colony of inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
By above three bacterium colonies that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultative any type of detesting in bacteria, severe bacteria or the non-bacterial infection.
Method such as embodiment 5 that its sample bacterium kind for the treatment of cultivation is identified.Then the bacterial classification that obtains is further carried out biochemical identification.
Embodiment 9: cultural method and this method interior with traditional anaerobic jar, the anaerobism culture bag are carried out parallel control experiment cultivation.
The example explanation
| Sample character | The example number | Needing to support environment cultivates | Detest and support the environment cultivation | The long bacterium example of symbiosis number | ||||||
| The inventive method | Detest and support bag | Detest and support jar | ||||||||
| Severe bacteria | Facultatively detest bacteria | Detest bacteria | Facultatively detest bacteria | Detest bacteria | Facultatively detest bacteria | Detest bacteria | Facultatively detest bacteria | |||
| Extract | 16 | ?8 | ?2 | ?6 | ?2 | ?5 | ?2 | ?2 | ?2 | Severe bacteria (8 example) |
| Facultatively detest bacteria (2 example) | ||||||||||
| Detest bacteria (6 example) | ||||||||||
| Secretory product | 9 | ?5 | ?1 | ?2 | ?1 | ?1 | ?1 | ?0 | ?1 | Severe bacteria (5 example) |
| Facultatively detest bacteria (1 example) | ||||||||||
| Detest bacteria (2 example) | ||||||||||
| No bacterial growth (1 example) | ||||||||||
| Ascites pleural fluid | 4 | ?1 | ?0 | ?1 | ?0 | ?0 | ?0 | ?0 | ?0 | Severe bacteria (1 example) |
| Facultatively detest bacteria (0 example) | ||||||||||
| Detest bacteria (1 example) | ||||||||||
| No bacterial growth (2 example) | ||||||||||
| Urine | 5 | ?2 | ?1 | ?1 | ?1 | ?0 | ?1 | ?0 | ?0 | Severe bacteria (2 example) |
| Facultatively detest bacteria (1 example) | ||||||||||
| Detest bacteria (1 example) | ||||||||||
| No bacterial growth (1 example) | ||||||||||
| Phlegm | 10 | ?4 | ?2 | ?2 | ?2 | ?2 | ?2 | ?1 | ?2 | Severe bacteria (4 example) |
| Facultatively detest bacteria (2 example) | ||||||||||
| Detest bacteria (2 example) | ||||||||||
| No bacterial growth (2 example) | ||||||||||
| Cerebrospinal fluid | 3 | ?0 | ?1 | ?0 | ?0 | ?0 | ?0 | ?0 | ?0 | Detest bacteria (1 example) |
| No bacterial growth (2 example) | ||||||||||
| Blood | 5 | 1 | 1 | 0 | 1 | 0 | 1 | 0 | 0 | Severe bacteria (1 example) |
| Facultatively detest bacteria (1 example) | ||||||||||
| No bacterial growth (3 example) | ||||||||||
Annotate: wherein, the severe bacteria of turning out has streptococcus aureus, staphylococcus epidermidis, alpha streptococcus, beta streptococcus, proteus, klepsiella pneumoniae genus etc.That turns out facultatively detests bacteria and streptococcus pneumoniae is arranged, Escherichia, staphylococcus etc.The bacteria of detesting of turning out has Bacteroides, Fusobacterium, bifid bar genus etc.Drug sensitive test slightly.
Cultivating the total routine number of all kinds of samples altogether by last table explanation is 52 examples, does not wherein have bacterial growth 11 examples; Turn out severe oxygen bacterium 21 examples, facultative anaerobe 8 examples are turned out anerobe 12 examples with present method, turn out anerobe 8 examples with anaerobism bag method, turn out anerobe 3 examples altogether with the anaerobism pot process.Thereby the anerobe positive rate height that adopts present method to cultivate as can be known, reliable results, method is easy, and the anaerobic bacteria culture, the expense that are fit to large, medium and small Microbiological Lab be low, simple and convenient and easy to study, is convenient to penetration and promotion.Method with biological oxygen consumption is created oxygen-free environment, replaces the method for physics, chemistry.Very big effect is played in the cultivation that popularizes anerobe.Utilize severe oxygen bacterium in the growth and breeding process with microenvironment in oxygen depletion produce a large amount of CO
2, cause anaerobic environment, help the growth and breeding of anerobe.Along with microbiological development, serial anaerobic bacteria culture relevant item is brought in constant renewal in, and the economical load of having avoided existing cultural method to cause to patient priorly causes difficulty and loss for clinical diagnosis, treatment.And novel method is simple, and is economical and practical, is suitable for hospitals at different levels, R﹠D institution, and experiment is accurate, reliable, suitability is strong.
The sample for the treatment of cultivation by this method can be cultivated by anaerobism simultaneously, thereby the sample that can treat cultivation is more accurately judged four kinds of situations that this sample may belong to: no bacterial growth, severe bacteria infect, detest the bacteria infection, facultative severe bacteria infects, its reliable results, method is easy, the kind that is fit to large, medium and small Microbiological Lab is identified, expense is low, simple and convenient and easy to study, is convenient to penetration and promotion.It has extremely practical meaning and value for clinical medicine.
Embodiment 10:
Adopt method of the present invention, be example to cultivate extract: its used vessel can adopt the device of the anaerobic bacteria culture of embodiment 1.
The first step, extract that will be to be cultivated are cultivated by embodiment 5 methods, the severe oxygen bacterium that wherein inoculates on the solid nutrient medium is intestinal bacteria, substratum with two kinds of different conditions: promptly in the device of anaerobic bacteria culture, carry out the substratum that the substratum cultivated under the oxygen-free environment and aerobic environment are cultivated, put into after 37 ℃ of constant incubators spend the night, found that, aerobic is cultivated no bacterial growth, and on the solid nutrient medium in the device that this anaerobism is cultivated the about 2mm of diameter is arranged, circular, dimpling, canescence, smooth surface, neat in edge, anhemolytic bacterium colony generates.The smear gramstaining, microscopic examination is found different in size, the blunt round and dense gram negative bacillus that dyes in two ends.Preliminary evaluation belongs to for the Gram-negative anaerobic bacillus(cillus anaerobicus).
Second step:
(1), the anerobe that the first step is cultivated out is seeded in respectively in glucose, lactose, maltose, glycosides dew alcohol, sucrose, pectinose, wood sugar, rhamnosyl, trehalose, salicin, Vitamin C2, liquefy gelatin, indole, catalase test and the 20% biliary micro-reaction assessor, above micro-reaction assessor is placed in the lower pot 2 of device of embodiment 1 a described new anaerobic bacteria culture, inoculates intestinal bacteria on the solid medium in the upper pot of this device.With the identical screwing hermetic of upper and lower part vessel, with isolate from outer air;
(2), adopt the used method of embodiment 5, the anerobe that the first step is cultivated out is evenly coated on the solid nutrient medium of lower pot of device of another new anaerobic bacteria culture, is scribbling the drug sensitive test paper that sticks penicillin, gentamicin, vancomycin, kantlex, erythromycin and Rifampin on the substratum of this anerobe respectively again.Solid medium inoculation intestinal bacteria in the upper pot of the device of this anaerobic bacteria culture.With the identical screwing hermetic of upper and lower part vessel, with isolate from outer air;
The device of two anaerobic bacteria culture that (1) and (2) is used is put into 37 ℃ of constant incubators together and is spent the night.Found that:
Drug sensitive test; This anerobe is to penicillin, gentamicin, vancomycin and kantlex resistance; Erythromycin and Rifampin sensitivity.
Biochemical identification: this anerobe is positive at the bile of glucose, lactose, maltose, sucrose, wood sugar, Vitamin C2, liquefy gelatin and 20%.Glycosides is revealed the reaction that is negative of alcohol, pectinose, rhamnosyl, trehalose, salicin, indole, catalase test.
Conclusion: identify that by above this anerobe is for being the bacteroides fragilis group.
Claims (13)
1, a kind of device of novel anaerobic bacteria culture, it is characterized in that, it is by last, the vessel that coincide mutually constitute down, periphery in arbitrary vessel of described upper pot or lower pot is provided with the isolation fastener that forms a circle, and the height of described isolation fastener is lower than the height of place vessel, form an interval between the outer wall of described isolation fastener and corresponding vessel, bottom in described interval is provided with Gask-O-Seal, the top edge of described another vessel is buckled on the Gask-O-Seal of the described vessel that are provided with Gask-O-Seal, described on, lower pot adopts screw thread or clamping to connect.
2, the device of a kind of novel anaerobic bacteria culture according to claim 1 is characterized in that, the bottom shape of described upper and lower part vessel is circle, ellipse, tetragon, Polygons or abnormity.
3, the device of a kind of novel anaerobic bacteria culture according to claim 1 is characterized in that, described isolation fastener be shaped as circle, ellipse, tetragon, Polygons or abnormity.
4, according to the device of claim 1,2 or 3 described a kind of novel anaerobic bacteria culture, it is characterized in that, the edge, outer ring of described Gask-O-Seal is consistent with the bottom shape of the described vessel that are provided with Gask-O-Seal, and the inner ring edge of described Gask-O-Seal is consistent with the shape of described isolation fastener.
5, the device of a kind of novel anaerobic bacteria culture according to claim 1 is characterized in that, the height of described upper and lower part vessel is 5-25mm, and the floorage of described upper and lower part vessel is 15cm
2-140cm
2, and be lower than the height of described another vessel at the height that is provided with the vessel of isolating fastener and Gask-O-Seal.
6, the device of a kind of novel anaerobic bacteria culture according to claim 1, it is characterized in that, be provided with the division board that encloses into a circle in vessel in described upper pot or lower pot or two vessel, described division board is divided into two intervals with the place vessel---interior interval and outer interval, the interval is the interval in the described division board circle in described, described outer interval is the outer interval of described division board circle, and the height of described division board will be lower than the height of its place vessel.
7, according to the device of claim 1,2,3,4 or 5 described a kind of novel anaerobic bacteria culture, it is characterized in that, in any vessel of described upper pot or lower pot, be provided with dividing plate, described dividing plate is divided into isolated two intervals mutually with its described vessel, and described dividing plate is lower than the height of place vessel.
8, the method for a kind of novel anaerobic bacteria culture according to claim 1 is characterized in that, includes following steps:
(1), at first prepares material---sample to be cultivated;
(2), step (1) gained sample being carried out oxygen-free environment in the device of described novel anaerobic bacteria culture cultivates;
The solid medium that has soon prepared places the vessel of the device of described novel anaerobic bacteria culture, cool off, solidify, the known severe bacteria of inoculation in filling vessel of described solid medium, inoculation sample to be cultivated in filling another vessel of described solid medium;
Or put into the solid medium that has prepared in the interior interval of the described upper and lower vessel in the device of described novel anaerobic bacteria culture, cool off, solidify, the known severe bacteria of inoculation on the interval solid medium in vessel is inoculated sample to be cultivated on the interval solid medium in described another vessel;
Or in the interior interval of arbitrary vessel of described upper pot in the device of described novel anaerobic bacteria culture or lower pot and outside the interval put into the solid medium that has prepared, cool off, solidify, the known severe bacteria of inoculation on the solid medium in interval in described, inoculation sample to be cultivated on the solid medium in interval outside described;
Or arbitrary vessel of the described upper and lower vessel in the device of described novel anaerobic bacteria culture, by putting into the solid medium that has prepared respectively in mutual isolated two intervals that dividing plate separated, cool off, solidify, the known severe bacteria of inoculation on the interval solid medium in the described part that separates by described dividing plate, inoculation sample to be cultivated on the interval solid medium in the described another part that separates by described dividing plate;
Described upper pot and described lower pot screwing hermetic are isolated with extraneous air, and the spacing between the described solid medium in wherein said upper pot and the described lower pot is at least greater than 5mm; Then the device of described novel anaerobic bacteria culture is put into 37 degrees centigrade of constant incubators and cultivated 10-18h, observe the growth of the bacterium of described inoculation sample to be cultivated;
(3), simultaneously step (1) gained sample is cultivated under common aerobic environment;
(4), by above three bacterial colonys that step observation post turns out, judged material to be cultivated---sample is to detest bacteria, facultative any type of detesting bacteria, severe bacteria or not having bacterial growth.
9, the method for a kind of novel anaerobic bacteria culture according to claim 8 is characterized in that, step (4) is to judge by the following method:
(A), sample described to be cultivated carried out oxygen-free environment cultivate in the device of novel anaerobic bacteria culture, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is severe bacteria, or does not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may be for detesting bacteria or facultatively detesting bacteria;
(B), sample described to be cultivated is cultivated on the common substratum that has under the foster environment, judge by the situation of bacterial growth:
(a), as there not being bacterial growth, sample then described to be cultivated is for detesting bacteria, or do not have bacterial growth;
(b), if any bacterial growth, sample then described to be cultivated may or facultatively be detested bacteria for severe bacteria;
Information summary by above-mentioned steps (A), (B) bacterial growth obtains four kinds of detected results,
(I), by (A, a) and (B, a) to obtain the result be no bacterial growth;
(II), by (A, a) and (B, b) to obtain the result be severe bacteria, promptly this sample is that severe bacteria infects;
(III), by (A, b) with (B, a) obtain the result and detest bacteria, promptly this sample infects for detesting bacteria;
(IV), to obtain the result by (A, b) and (B, b) be facultative severe bacteria, promptly this sample is that facultative severe bacteria infects.
10, the method for a kind of novel anaerobic bacteria culture according to claim 8 is characterized in that, sample described to be cultivated is human body or animal body excretory extract, phlegm, ascites pleural fluid, secretory product, cerebrospinal fluid, blood, urine etc.
11, the method for a kind of novel anaerobic bacteria culture according to claim 8 is characterized in that, described solid medium is that solid-based basal culture medium, solid nutrient medium, solid are selected any of substratum, solid differential medium.
12, the method for a kind of novel anaerobic bacteria culture according to claim 8 is characterized in that, this method can be used as the drug sensitivity test of detesting bacteria.
13, the method for a kind of novel anaerobic bacteria culture according to claim 8 is characterized in that, the described thickness of putting into the off-the-shelf solid medium of upper and lower part vessel is at least 3mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200410090290XA CN1295322C (en) | 2004-11-04 | 2004-11-04 | Anaerobic bacteria culturing installation and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200410090290XA CN1295322C (en) | 2004-11-04 | 2004-11-04 | Anaerobic bacteria culturing installation and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1635109A true CN1635109A (en) | 2005-07-06 |
| CN1295322C CN1295322C (en) | 2007-01-17 |
Family
ID=34847594
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200410090290XA Expired - Fee Related CN1295322C (en) | 2004-11-04 | 2004-11-04 | Anaerobic bacteria culturing installation and method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1295322C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242074A (en) * | 2011-04-12 | 2011-11-16 | 叶春 | Screen method for facultative anaerobe, and apparatus thereof |
| CN109576130A (en) * | 2018-12-18 | 2019-04-05 | 东北农业大学 | Artificially manufacture the device that micro- oxygen environment purifies micro- oxygen bacterium |
| CN110072988A (en) * | 2016-10-28 | 2019-07-30 | 国立大学法人京都大学 | The co-culture device and co-culture method of the bacteriums such as Anaerobic Bacteria and epithelial cell |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IE56078B1 (en) * | 1982-09-30 | 1991-04-10 | Wadley Technologies Inc | Detection of microbial pathogens |
| CN2055136U (en) * | 1989-10-04 | 1990-03-28 | 蚌埠医学院 | Culture device for anaerobic bacteria |
-
2004
- 2004-11-04 CN CNB200410090290XA patent/CN1295322C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242074A (en) * | 2011-04-12 | 2011-11-16 | 叶春 | Screen method for facultative anaerobe, and apparatus thereof |
| CN102242074B (en) * | 2011-04-12 | 2014-08-06 | 叶春 | Screen method for facultative anaerobe |
| CN110072988A (en) * | 2016-10-28 | 2019-07-30 | 国立大学法人京都大学 | The co-culture device and co-culture method of the bacteriums such as Anaerobic Bacteria and epithelial cell |
| CN109576130A (en) * | 2018-12-18 | 2019-04-05 | 东北农业大学 | Artificially manufacture the device that micro- oxygen environment purifies micro- oxygen bacterium |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1295322C (en) | 2007-01-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN87106707A (en) | The equipment of microorganism culturing and detection and method | |
| CN101067119A (en) | Fermenting lactobacillus CMS II002 and its application | |
| CN101032281A (en) | Fodder additive having immunologic enhancement in growing and preparation method thereof | |
| CN1207382C (en) | Lactob.plantarum ST-III strain and application in regulating blood fat | |
| CN1295322C (en) | Anaerobic bacteria culturing installation and method | |
| CN1274803C (en) | Model of mouth cavity biomembrane and method for forming mouth cavity biomembrane | |
| CN106868094A (en) | The method for quick of antibiotic residue in a kind of raw milk | |
| CN1858200A (en) | Oral biological film dynamic model device and its oral biological film forming method | |
| CN1763173A (en) | Hematopoietic stem cell culturing apparatus and method | |
| CN100338206C (en) | Microbial culture medium and use thereof | |
| CN200952006Y (en) | Oral cavity biological membrane dynamic model device | |
| CN218026056U (en) | A bacterial culture apparatus for quick purification treatment of sewage | |
| CN1317385C (en) | Cheese lactobacillus Bd-II strain and use for decreasing blood fat | |
| CN102676407A (en) | Microzyme for degrading residual lincomycin in fermented leaves and application thereof | |
| CN204162724U (en) | A kind of perfusion cap cultivated for biological sample filling type | |
| CN1858238A (en) | Quick tubercle myco-bacillus culture medium | |
| CN111040953B (en) | Method for rapidly screening variant microorganisms based on PET imaging system | |
| CN206538435U (en) | A kind of multi-functional medical test microbial culture garden | |
| CN213537905U (en) | Bioreactor (mixing) shaft rapid disassembly structure | |
| CN1934268A (en) | Quality ensurement system for detecting microorganisms | |
| CN212128203U (en) | Culture device suitable for research of interaction between cultures through gas communication | |
| CN1556217A (en) | Method of fast screening pathogenic microorgnnism sensitive medicine | |
| CN216585038U (en) | Bacterial biofilm drug resistance detection device | |
| CN202543220U (en) | High-throughput cell culture incubator | |
| CN105441326A (en) | Perfusion cover for biological sample perfusion culture and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CB03 | Change of inventor or designer information |
Inventor after: Tang Zhengliang Inventor after: Wang Huiyong Inventor after: Wang Mao Inventor before: Wang Huiyong Inventor before: Wang Mao |
|
| COR | Change of bibliographic data | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070117 Termination date: 20211104 |