CN1213149C - Counting card for bacteria, funguses and caliform floras and its preparing process - Google Patents
Counting card for bacteria, funguses and caliform floras and its preparing process Download PDFInfo
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- CN1213149C CN1213149C CN 01107661 CN01107661A CN1213149C CN 1213149 C CN1213149 C CN 1213149C CN 01107661 CN01107661 CN 01107661 CN 01107661 A CN01107661 A CN 01107661A CN 1213149 C CN1213149 C CN 1213149C
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- counting
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- underframe
- filter paper
- sterilization
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Abstract
The present invention provides a counting card for bacteria, fungi and coliform, and a preparation method of the counting card. The counting card is composed of a counting card box and an inner counting card which is arranged on the bottom of the counting card box, wherein the inner counting card is composed of a lower frame, an upper frame and a counting sheet which is clamped between the upper frame and the lower frame; the counting sheet which is composed of microporous filtering films of upper layer material and filter paper of lower layer material adsorbing a sterilizing liquid culture medium is concave downwards to form a vessel shape with a shallow flat bottom. The counting card can be used for quickly measuring the total number of bacteria, the total number of fungi and the number of coliform in food, beverages, mineral water, drinking purified water, tap water and medicines, and the detection sensitivity is greatly improved.
Description
The present invention relates to microorganism detection device and preparation method thereof.
At food, beverage, mineral water, drink in the check of pure water, makeup, medicine and feed, total plate count, total number of fungi and coliform quantity are crucial limit detection indexs, than chemical substance, microorganism is more active, have more uncontrollability and foresight, thereby cause that always showing great attention to of people, Recent study develop the then heat subject in biological technical field especially of this type of rapid detection product.
Early eighties, Minnesota Mining and Manufacturing Company at first is engaged in the research of this respect, produce Petrifilm series bacterium, fungi tally and coliform counting film, only need during use directly to be coated on sample or its diluent on the film, count the bacterium colony that has color after cultivating and get final product, operation is very easy.Reports such as Kinght MT in 1997, the yeast of Minnesota Mining and Manufacturing Company and mold count plate and traditional method do not have significant difference.After 3M company, the close row pool of Japanese Y company, the various dry plate products that also release one after another such as U.S. Drew chemical company and Finland Orion Diagnostica factory.
The domestic microorganism detection dry plate product that has formed patent or product has bacteriologic test card in the serial water, total number of bacterial colony to detect glued membrane, the coliform quick test scraps of paper and the mould check scraps of paper etc. at present, greatly promoted the development of domestic microbial testing technology, and obtain to a certain degree application in practice, and tableware coliform paper disk method has also been listed national standard in.
In mentioned microorganism check goods, though total number of bacterial colony detection glued membrane sensing range is wide, diffustivity is strong thereon for many bacterium, can not form bacterium colony well, therefore can have a strong impact on result's reliability; Bacterium, mould and the coliform test scraps of paper mostly are to adopt filter paper as carrier, because bacterium and mould spores and yeast cell are all less, therefore cause them to grow at the filter paper different depths easily, and can't count; In addition, some tool flagellums or the strong bacterium of mobility can be come along the diffusion of the hole of filter paper fibre, make bacterium colony smudgy, sometimes even diffuse to the monoblock scraps of paper and can't count; And,, prepare comparatively loaded down with trivial details because the levels of check card needs to glue connection with Chemical Felter by the bacteriologic test card that millipore filtration and the sticking connection of nutrition liner form.Each above-mentioned based article all needs sterile petri dish to cooperate in use, uses inconvenience, is unfavorable for carrying out quick test.
The objective of the invention is to overcome the obvious deficiency in the present microorganism dry plate testing product, total plate count, total number of fungi and coliform quantity in direct tracer liquid or the semi-solid sample can be provided, and accuracy and traditional method do not have novel bacteria, fungi and the coliform numbered card of significant difference, be used for rapid determination, and improve detection sensitivity greatly food, beverage, mineral water, the total plate count of drinking pure water, tap water and medicine, total number of fungi and coliform quantity.
The bacterium that is provided among the present invention, fungi and coliform numbered card, by the counting cartridge be positioned over and block two portions in the counting of this counting cartridge bottom and form, the bottom of said counting cartridge is smooth, bottom shape matches with the interior card of counting, card is by underframe in the said counting, upper frame and being clipped in, count slice between the underframe constitutes, the inwall of said underframe has the bead of a staged along bottom periphery, said counting horse is on the ladder bead of said underframe, along being recessed into shallow flat ware shape under the preglabellar field of this ladder bead, so that placement testing liquid sample does not wherein outflow is degree, upper frame presses solidly the edge clamping of count slice on the bead of underframe, said count slice is by last, following two layers of material constitutes, its subsurface material is a filter paper, be adsorbed with the liquid nutrient medium after the sterilization on the filter paper, upper layer of material is a millipore filtration.
Can print the counting lattice on the said count slice, so that enumeration to growing.
It is the millipore filtration of 0.2～1.0um that the upper layer of material of said count slice can adopt the aperture.
The height of the underframe of said numbered card is generally 3～8mm, and the ladder lug height of this underframe is 1～5mm.
The upper frame and the underframe of said counting cartridge and numbered card can be made by transparent plastics.
Said counting cartridge can be designed to different shape as required, is preferably circular or square.
Liquid nutrient medium on the above-mentioned mentioned filter paper that is adsorbed on count slice, both can adopt general liquid nutrient medium, also can adopt and have the opposing sanitas, the liquid nutrient medium of sterilizing agent or ozone jamming performance, for example produce by Guangdong Microbes Inst subordinate Huankai Microbiological Science ﹠ Technolgy Co., Guangdong and public offering have an opposing sanitas, the anti-interference liquid culture based articles of sterilizing agent or ozone jamming performance, ten two prescriptions listed as table 1, these liquid nutrient mediums have the opposing sanitas respectively, sterilizing agent or ozone interferential performance, contain anti-sanitas in detection, during the sample of interfering substance such as sterilizing agent or ozone, compare with common numbered card, can improve the sensitivity of detection greatly.Table 1 is 12 different ingredients liquid nutrient mediums and consumption and the made numbered card of producing with Huankai Microbiological Science ﹠ Technolgy Co., Guangdong:
The substratum of the various numbered cards of table 1 and consumption
| The numbered card title | The substratum that uses | Consumption (g/L) |
| The bacterial count card | Nutrient broth medium | 22 -24 |
| Anti-anticorrosion formulation bacterial count card | Anti-anticorrosion formulation bacterial liquid full-page proof detects substratum | 22 -28 |
| Anti-sterilization formulation bacterial count card | Anti-sterilization formulation bacterial liquid full-page proof detects substratum | 20 -24 |
| Anti-ozone type bacterial count card | Anti-ozone type bacterial liquid full-page proof detects substratum | 19 |
-23
14
Fungi numbered card mould liquid nutrient medium
-18
Anti-anticorrosion formulation fungi liquid full-page proof detects cultivates 21
Anti-anticorrosion formulation fungi numbered card
Base-25
Anti-sterilization formulation fungi liquid full-page proof detects cultivates 17
Anti-sterilization formulation fungi numbered card
Base-21
16
The anti-ozone type fungi liquid of anti-ozone type fungi numbered card full-page proof detects substratum
-20
32
Coliform numbered card lactose cholate fermention medium
-40
The anti-anticorrosion formulation coliform tracer liquid of anti-anticorrosion formulation coliform meter cultivates 36
Number Ka Ji-44
The anti-sterilization of anti-sterilization formulation coliform meter formulation coliform tracer liquid cultivates 33
Number Ka Ji-41
Anti-ozone type coliform counting 32
Anti-ozone type coliform tracer liquid substratum
Card-40
Be the concrete step of preparation process of bacterium of the present invention, fungi and coliform numbered card below:
(1), by the upper frame and the underframe of card in size preparation counting cartridge and the counting, millipore filtration and the filter paper that constitutes the numeration sheet is cut into the size that matches with underframe;
(2), sterilising treatment:
A, with cobalt-60 or oxyethane will count cartridge and the counting in the card upper frame and underframe carry out sterilising treatment;
B, filter paper and millipore filtration are carried out sterilising treatment with cobalt-60 or high-temperature steam;
C, liquid nutrient medium is sterilized with high-temperature steam or membrane filtration;
(3), by sterilization filter paper institute can the adsorption liquid scale of construction 1-5 doubly measure the liquid nutrient medium of drawing after the above-mentioned sterilization, the filter paper of sterilizing is joined in this culture medium after sterilization adsorb then, or culture medium after sterilization is joined in the filter paper of sterilization;
(4), take out the filter paper that fully adsorbs substratum, place on the ladder bead of underframe, form count slice at the millipore filtration that attaches sterilization on the filter paper again, press-fiting upper frame then is fixed on count slice on the ladder bead of underframe, again count slice is depressed to shallow flat ware shape along the preglabellar field of the ladder bead of said underframe, card in the counting;
(5), holding 35-100 ℃ baking oven inner drying 0.5-3.0 hour in the counting;
(6), the bottom that is placed in the counting cartridge in the exsiccant counting, cover plastic box cover, promptly make described bacterium, fungi and coliform numbered card.
The present invention has solved the subject matter of existing bacterium, fungi and the existence of coliform test dry plate product both at home and abroad preferably, numbered card all adopt certain concavity arranged around the bottom mould the box integrated apparatus, needn't cooperate with sterile petri dish, can make things convenient for application of sample and carry out rapid detection, make bacterium and coliform reduce the bacterium colony diffusion, and make fungal growth good, be convenient to counting.In addition, can be made into anti-anticorrosion formulation, anti-sterilization formulation and anti-ozone type numbered card to eliminate sanitas Sorbic Acid and potassium sorbate, phenylformic acid and Sodium Benzoate respectively, the interference (table 2) of sterilizing agent dioxide peroxide, Peracetic Acid, clorox and hydrogen peroxide and ozone and chlorine residue, when detection contains the sample of interfering substance, compare with common numbered card, can improve the sensitivity of detection greatly.
Interfering substance and concentration thereof that the anti-interference numbered card of table 2 can be eliminated
The dried chaff interference that can eliminate that can eliminate
The numbered card title
Disturb the concentration of material matter
Phenylformic acid, benzene first
Acid sodium
Anti-anticorrosion formulation numbered card 2.0g/L (kg)
Sorbic Acid, sorb
Acid potassium
Dioxide peroxide 150mg/L
Peracetic Acid 400mg/L
Anti-sterilization formulation numbered card
Clorox 700mg/L
Hydrogen peroxide 180mg/L
Ozone
Antiozonidate type numbered card 10.0mg/L
Chlorine residue
Fig. 1 is the front view of one embodiment of the invention;
Fig. 2 is the side-view of Fig. 1;
Fig. 3 is that the A-A of Fig. 1 is to sectional view;
Fig. 4 is the front view of the interior card of counting among Fig. 1;
Fig. 5 is that the B-B of Fig. 4 is to sectional view.
Below in conjunction with accompanying drawing embodiment of the present invention is described in further detail:
Embodiment one
This bacterium, the anti-interference numbered card of fungi and coliform is by counting cartridge 1 and be positioned over and block 2 two portions in the counting of this counting cartridge bottom and form, counting cartridge 1 is made of box body 11 and lid 12, adorn card 2 in two identical square countings in the counting cartridge 1, card 2 comprises a quadrate underframe 22 and a quadrate upper frame 23 in the counting, the count slice 21 that forms shallow flat ware shape is clipped in, between the underframe, (23 edge clamping with count slice 21 press solidly on the ladder bead of underframe 22 upper ledge, count slice 21 is by last, following two layers of material constitutes, its subsurface material is a filter paper, be adsorbed with the liquid nutrient medium after the sterilization on the filter paper, upper layer of material is the aperture millipore filtration, is printed with the counting grid on it.
The preparation method of this numbered card is as follows:
1, select for use polystyrene as counting cartridge 1 and numbered card 2 upper and lower frame materials, the size of counting cartridge is 88 * 44 * 15mm, the wall thickness of this counting cartridge is 2mm, about being divided into two every, the underframe 22 of card and the size of upper frame 23 are respectively 40 * 40 * 5mm and 35 * 35 * 2mm in the counting, the ladder lug height of underframe is 2mm, and upper frame can overlap in the inside casing wall that is sandwiched in underframe, and designing mould is made card plastic frame in plastics counting cartridge and the counting in view of the above;
2, adopting cobalt-60 to carry out sterilization by blocking good filter paper of the big cabinet of underframe and millipore filtration and plastics counting cartridge and plastic frame in the counting.
3, all kinds of liquid nutrient mediums that do not contain chlorination triphenylmethyl tetrazole (TTC) are placed 121 ℃, 0.1MPa sterilized 15-20 minute down; TTC solution then adopts 0.2 μ m filtering with microporous membrane sterilization, treat the substratum cooling after, add before using and wherein get final product.
4, under aseptic condition, draw liquid nutrient medium after the above-mentioned sterilizations by sterilization filter paper institute can the adsorption liquid scale of construction 3 times, then the filter paper of sterilizing is placed in one, adsorb after 10 minutes, after placing on the interior card of the opened counting frame of plastic lower frame, on filter paper, place filter membrane, and the upper ledge that closes.
5, put count slice and after 2.0 hours, take out the bottom that the exsiccant count slice places the sterilization plastics casing in 70 ℃ baking oven inner drying, plastic cover closes.
6, after being put heat-shrinkable film, packaged numbered card carries out thermal contraction, the outer packing box of packing into then.
It is as follows to adopt above-mentioned numbered card to carry out the method for counting of bacterium, fungi and coliform:
1, dissolving in the solid sample adding sterile saline or making semi-solid sample, liquid sample can directly detect, and need not sample is handled.
2, behind the numbered card of opening embodiment one production on the Bechtop, draw the 1mL sample, join the central authorities of numbered card film, the plastic box cover that closes then treats after 5 minutes that the liquid on the film is all blotted by the filter paper of lower floor, is inverted numbered card.
3, bacterium and the holding of coliform counting were cultivated 24-36 hour for 35-37 ℃, the holding of fungi counting is cultivated after 36-48 hour for 30-32 ℃, observes counting.
4, on bacterium and coliform numbered card, show red and have the quantity of obvious projection to be respectively bacterium and coliform group count.There are mildew or thread quantity then to be fungi count on the fungi numbered card, have the bacteroid projection colony number of class then to be yeast count.
5, the counting bacterium number that is fit to of numbered card is less than 200cfu/mL, then need adopt after the stroke-physiological saline solution dilution counting again as surpassing.
Claims (7)
1. bacterium, fungi and coliform numbered card, it is characterized in that forming by counting cartridge (1) and interior (2) two portions that block of counting of being positioned over this counting cartridge bottom, the bottom of said counting cartridge (1) is smooth, bottom shape matches with the interior card of counting (2), card (2) is by underframe (22) in the said counting, upper frame (23) and being clipped in, count slice between the underframe (21) constitutes, the inwall of said underframe (22) has the bead of a staged along bottom periphery, said count slice (21) frame is on the ladder bead of said underframe (22), along being recessed into shallow flat ware shape under the preglabellar field of this ladder bead, upper frame (23) presses solidly the edge clamping of count slice (21) on the bead of underframe (22), said count slice (21) is by last, following two layers of material constitutes, its subsurface material is a filter paper, be adsorbed with the liquid nutrient medium after the sterilization on the filter paper, upper layer of material is a millipore filtration.
2. numbered card according to claim 1 is characterized in that being printed on the counting lattice on the said count slice (21).
3. numbered card according to claim 1, the aperture that it is characterized in that the upper layer of material millipore filtration of said count slice (21) is 0.2～1.0um.
4. numbered card according to claim 1 is characterized in that the height of the underframe (22) of card (2) in the said counting is 3～8mm, and the ladder lug height of this underframe is 1～5mm.
5. numbered card according to claim 1 is characterized in that the upper frame (23) and the underframe (22) of card (2) in said counting cartridge (1) and the counting made by transparent plastics.
6. numbered card according to claim 1 is characterized in that said counting cartridge (1) is for circular or square.
7. the preparation method of the described bacterium of claim 1, fungi and coliform numbered card is characterized in that preparation process is as follows:
(1), by upper frame (23) and the underframe (22) of card (2) in size preparation counting cartridge (1) and the counting, the millipore filtration and the filter paper of formation count slice is cut into the size that matches with underframe;
(2), sterilising treatment:
A, with cobalt-60 or oxyethane will count cartridge (1) and the counting in the card (2) upper frame (23) and underframe (22) carry out sterilising treatment;
B, filter paper and millipore filtration are carried out sterilising treatment with cobalt-60 or high-temperature steam;
C, liquid nutrient medium is sterilized with high-temperature steam or membrane filtration;
(3), by sterilization filter paper institute can the adsorption liquid scale of construction 1-5 doubly measure the liquid nutrient medium of drawing after the above-mentioned sterilization, the filter paper of sterilizing is joined in this culture medium after sterilization adsorb then, or culture medium after sterilization is joined in the filter paper of sterilization;
(4), take out the filter paper that fully adsorbs substratum, place on the ladder bead of underframe (22), form count slice (21) at the millipore filtration that attaches sterilization on the filter paper again, press-fiting upper frame (23) then is fixed on count slice on the ladder bead of underframe, again count slice (21) is depressed to shallow flat ware shape along the preglabellar field of the ladder bead of said underframe, card (2) in the counting;
(5), card (2) in the counting is put 35-100 ℃ baking oven inner drying 0.5-3.0 hour;
(6), card (2) in the exsiccant counting is placed the bottom of counting cartridge, cover plastic box cover, promptly make described bacterium, fungi and coliform numbered card.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01107661 CN1213149C (en) | 2001-03-23 | 2001-03-23 | Counting card for bacteria, funguses and caliform floras and its preparing process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01107661 CN1213149C (en) | 2001-03-23 | 2001-03-23 | Counting card for bacteria, funguses and caliform floras and its preparing process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1376800A CN1376800A (en) | 2002-10-30 |
| CN1213149C true CN1213149C (en) | 2005-08-03 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01107661 Expired - Fee Related CN1213149C (en) | 2001-03-23 | 2001-03-23 | Counting card for bacteria, funguses and caliform floras and its preparing process |
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|---|---|---|---|---|
| CN101906384B (en) * | 2009-06-03 | 2013-03-27 | 中国科学院电子学研究所 | Instrument and method for bacterium flow count testing |
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| CN1376800A (en) | 2002-10-30 |
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