CN1051758A - Total number of bacterial colony detects glued membrane and preparation method - Google Patents
Total number of bacterial colony detects glued membrane and preparation method Download PDFInfo
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- CN1051758A CN1051758A CN 90108397 CN90108397A CN1051758A CN 1051758 A CN1051758 A CN 1051758A CN 90108397 CN90108397 CN 90108397 CN 90108397 A CN90108397 A CN 90108397A CN 1051758 A CN1051758 A CN 1051758A
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Abstract
A kind of total number of bacterial colony detects glued membrane and preparation method thereof, adopt the wide aperture glass fibre to do carrier, the nutritive medium that hot dipping is made in peptone, yeast extract, Zulkovsky starch, Gelucystine, gelatin, agar, meat extract, the immersion liquid of the liver heart etc., and then be dipped in the nutritive medium that glucose, blood digest broth are made, take out the back and spray triphenyltetrazolium chloride and toluylene red, oven drying at low temperature and making, this glued membrane can detect the total number of bacterial colony of food, makeup, tableware, body surface, water etc., and sensing range is wide, the detection operation is simple.
Description
The present invention relates to total number of bacterial colony and detect glued membrane and preparation method.
Total number of bacterial colony is to detect food, body surface, makeup, tableware and water etc. by an important indicator of bacterial contamination degree.General in the world plate tilt-pour process of the past, but material, complex operation are taken a lot of work, taken to this method, and the basic unit laboratory is difficult for promoting.In recent years both at home and abroad all in the fast simple detection means of research.A kind of colony determination card has been introduced by the U.S. in " the total number of bacterial colony specification sheets " that print off in April, 1989, can check the total number of bacterial colony of water and air.Contained in Japan's 89 years " technology lecture " " quantitatively bacteriologic test method summary " VOL17.No6.P536 has put down in writing a kind of ス Application Block method of checking total number of bacterial colony, can detect the pollution of finger, bed, wall.Domestic in 1986 under Academy of Medical Sciences expert instructs, Chongqing second health and epidemic prevention station has developed water and water sample sample total number of bacterial colony detector; Tianjin Environmental Protection Agency has developed the tap water check card, and these two kinds of products all are only limited to the check of water and water sample sample.
The detector of analogous products water from the research idea analysis, all is to utilize the habitual degerming filter membrane of microbiology to do carrier both at home and abroad, and the nutrition pad is put in lower floor, and the outside is overlapped with framed, utilizes degerming filter membrane aperture little, bacterium is blocked up stayed on the face.If utilize the degerming filter membrane to do carrier, because hole is too little, water molecules is by all very difficult, and water is checked in inevitable supply only.Can find out that from the above these prior art sensing ranges limit to, and the technology trouble, poor accuracy, cost height.
The object of the present invention is to provide a kind of still unsolved at present multi-functional total number of bacterial colony to detect glued membrane and preparation method both at home and abroad, simplify test step, enlarge sensing range.
Technical solution of the present invention is the preparation method that total number of bacterial colony detects glued membrane, follows these steps to make:
A, get peptone 20 gram, yeast extract 1 gram, 400 milliliters of Zulkovsky starch 3 grams, sodium-chlor 5 grams, dipotassium hydrogen phosphate 2.6 grams, Gelucystine 0.13 gram, gelatin 5 grams, agar 5 grams, meat extract, 100 milliliters of liver heart immersion liquid, adding distil water to 1000 milliliter, heating is dissolved and is transferred PH to 7.4, sterilization was the nutritive medium I in 15 minutes under 15 pounds of pressure
B, get glucose 3 gram and add in 200 milliliters of the blood digest broths and be the nutritive medium II.
C, wide aperture glass fiber paper are done carrier and are put into 90 ℃ of hot nutritive medium I and soak into, and then are dipped in 60 ℃ of nutritive medium II, and take out the back and spray triphenyltetrazolium chloride and toluylene red, oven drying at low temperature, pack is with cobalt
60The irradiation of 100～2,500,000 rad doses gets final product.
Method peptone of the present invention is got tryptone 5 gram, many peptones 10 grams, soya peptone 5 grams, the immersion liquid of the liver heart and is got 20 milliliters of liver infusions, 80 milliliters of heart immersion liquid.
Each bacterioid Different Nutrition of product analysis external environment of the present invention requires and has selected the nutrition agent peptone, Zulkovsky starch, and the immersion liquid of the liver heart, yeast, glucose have been selected weighting agent and sorbent material gelatin and agar.This is the basic composition of glued membrane, and it is attracted on the film bacterium to grow, and membrane permeability of making thus and good dispersity can be accepted the check of macrobead sample.Product of the present invention has adopted compound indicator triphenyltetrazolium chloride and toluylene red, and it guarantees under the general condition colourless, makes the bacterium colony colour developing when having only bacterial growth.Product of the present invention has been sought toughener Gelucystine, blood digest broth, the immersion liquid of the liver heart of indicator, and this toughener not only strengthens the colourity of indicator, simultaneously bacterial growth is had promoter action.The judgement of total number of bacterial colony must be carried out at 24 hours, and, a part of bacterium just showed on glued membrane after 24 hours gradually.Add the positive growth factor blood digest broth, this part bacterium just can be grown at 16 hours.The sodium-chlor and the dipotassium hydrogen phosphate of product of the present invention play shock absorption, guarantee that bacterial equilibrium is injury-free.
Total number of bacterial colony of the present invention detects glued membrane and has the following advantages:
1, the water flowing of this product glued membrane is good, sample can spread rapidly on face, this specific character makes the sample of band bits also can check, so be applicable to multiple sample, promptly all can detect, thereby scope of survey is than checked the examination and test of products amount of water big 20～30 times in the past to food, makeup, tableware, body surface, water etc.
2, compare with check card, Tianjin check card, the Chongqing detector of the U.S., easy to use, application of sample is accurate, and denomination is big, and (product of the U.S. has only 1.5cm
2, Tianjin product 7cm
2, Chongqing product 10cm
2, this product 48cm
2), can hold 2000 bacteriums and evenly grow.
3, compare with dull and stereotyped tilt-pour process, can remove purchasing of glass equipment from, store, scrub, sterilization, equipment are damaged consumes, dissolves pour plate, remove purchasing, store and making of culture medium raw material from, thus test step simplified, easy to use, and bacterium colony takes on a red color, be easy to identification, counting accurately can reduce eyes muscular fatigue again.
4, product of the present invention can use half a year in room temperature preservation, be convenient to transportation, can put half a year in the refrigerator after the detection, be convenient to superintendent office's periodic reinvestigation, it and the supporting use of coliform detection paper, ready-to-eat greatly, environmental sanitary inspection department is to the supervision and management of basic unit's work of cement labouratory quality, for realizing that product checks batch by batch qualified dispatching from the factory and creates conditions.
Further specify technical solution of the present invention in conjunction with example.Method of the present invention is operated according to the following steps:
1, the preparation of meat, the heart, liver infusion
Getting 1000 gram degrease hamburgers or the hearts, liver adds 2000 ml waters and boils after 15 hours at the refrigerator cold soaking and made in 1 hour.
2, the preparation of blood digest broth
Get 1000 milliliters of ox bloods, add distilled water and boiled 15 fens for 1000 milliliters, be cooled to 40 ℃ and add yellow soda ash accent pH value 8.5, add 60 milliliters of pancreatin again, under 37 ℃ of temperature, placed 48 hours, through peptone qualitative test feminine gender, stop digestion, filter, in refrigerator, preserve standby.
3, get tryptone 5 grams, many peptone 10 grams, soya peptone 5 grams, yeast extract 1 gram, Zulkovsky starch 3 grams, sodium-chlor 5 grams, dipotassium hydrogen phosphate 2.6 grams, Gelucystine 0.13 gram, gelatin 5 grams, agar 5 grams, 400 milliliters of meat extracts, 20 milliliters of liver infusions, 80 milliliters of heart immersion liquid, adding distil water to 1000 milliliter, heating is dissolved and is transferred PH to 7.4, and sterilization was the nutritive medium I in 15 minutes under 15 pounds of pressure.
4, get glucose 3 grams and add 200 milliliters of blood digest broths, be the nutritive medium II.
5, getting wide aperture glass fiber paper that Hangzhou, Zhejiang province city Xinhua Paper Making Mill produces does carrier and puts into 90 ℃ of hot nutritive medium I and soak into, and then be dipped in 60 ℃ of nutritive medium II, and take out the back and spray triphenyltetrazolium chloride and toluylene red, under 40 ℃ of temperature, dry, pack is with cobalt
60The irradiation of 100～2,500,000 rad doses gets final product.
The using method of product of the present invention: after sample was handled routinely, each extent of dilution respectively added 1 ml sample with two glued membranes, and take out sample infiltration back, puts under 37 ℃ of temperature condition and cultivated 18～24 hours, looks into the red bacterium colony on the glued membrane, routinely the report of method counting.
Application example
One, censorship unit: Harbin Sanitation and Antiepidemic Statin
Examination unit: Institute of Microorganism, Academia Sinica
Two, the censorship date: in August, 1988, in March, 1989
Three, censorship purpose:
(1) can all kinds of bacterium grow on glued membrane
(2) glued membrane method and plate tilt-pour process have not difference on the reflection colony counts.
Four, method:
(1) measure bacterial strain:
1,365 colon bacillus
2,89 streptococcus aureuses
3,31 proteus vulgaris
4,8821-1 bacillus licheniformis
5,8816 bacillus pumiluses
6,8840,8514 Pseudomonas aeruginosas
7,595 streptococcus faecium
8,8801 candiyeasts
9,396 carbon black aspergillus
(2) step:
1, bacteria suspension preparation: 24 hours slant culture of picking one little ring, evenly be suspended in 10 milliliters of stroke-physiological saline solution, with 100 times be diluted to 100,000 successively/.
2, cultivate: select suitable concn, get 1 milliliter respectively and add to glued membrane, cultivated 48 hours at 37 ℃.
3, counting: the red point on the scraps of paper is counted.
Five, result:
(1) growth test of all kinds of bacterium: glued membrane can be supported the growth of the mushroom of measuring, and sees table one for details.
The growth measurement of all kinds of bacterium of table one
| The bacterium name | Bacterial strain number | Growing state |
| Arthrobacterium Candida pseudomonas aeruginosa bacillus pumilus ETEC staphylococcus aureus carbon black aspergillus | 8801 2.1182 8514 8816 1.365 1.89 3.396 | + + + + + + + |
(2) comparison of glued membrane method and Plating: glued membrane method and Plating are variant slightly, see Table 2.
The comparison of table two glued membrane method and flat band method
| Bacterium number | Glued membrane (individual/1 ML) | Dull and stereotyped | ||||
| 10 5 | 10 6 | 10 7 | 10 5 | 10 6 | 10 7 | |
| 1.365 1.89 31 8821-1 1.595 8840 | 443345 133 240 767 | 302106 117385 26 53 | 27 | 560840 784 190 2880 | 161160 130760 135 25 | 11 |
Six, conclusion:
(1) bacterium of being measured, yeast and aspergillus all can be grown on glued membrane, and glued membrane nutrition can be supported multiple fungus grown, and range of application also can enlarge thereupon.
(2) as seen, glued membrane method counting is lower than Plating slightly, and the two counting error is on the same order of magnitude from table two.
(3) shown red colony edge is clear, even the distortion flora of commute diffusion also can see clearly single bacterium colony, is convenient to counting.
Claims (4)
1, a kind of total number of bacterial colony detects the preparation method of glued membrane, it is characterized in that adopting following method to make,
A, get peptone 20 gram, yeast extract 1 gram, Zulkovsky starch 3 grams, sodium-chlor 5 grams, dipotassium hydrogen phosphate 2.6 grams, Gelucystine 0.13 gram, gelatin 5 grams, agar 5 grams, 400 milliliters of meat extracts, 100 milliliters of liver heart immersion liquid, adding distil water to 1000 milliliter, heating is dissolved and is transferred PH to 7.4, sterilization was the nutritive medium I in 15 minutes under 15 pounds of pressure
B, get glucose 3 gram and add in 200 milliliters of the blood digest broths and be the nutritive medium II,
C, wide aperture glass fiber paper are done carrier and are put into 90 ℃ of hot nutritive medium I and soak into, and then are dipped in 60 ℃ of nutritive medium II, and take out the back and spray triphenyltetrazolium chloride and toluylene red, oven drying at low temperature, pack is with cobalt
60The irradiation of 100～2,500,000 rad doses gets final product.
2, the method for claim 1 is characterized in that peptone gets tryptone 5 gram, many peptones 10 grams, soya peptone 5 grams.
3, method as claimed in claim 1 or 2 is characterized in that the immersion liquid of the liver heart gets 20 milliliters of liver infusions, 80 milliliters of heart immersion liquid.
4, a kind of total number of bacterial colony detects glued membrane, it is characterized in that adopting the product of the method preparation of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN90108397A CN1039825C (en) | 1990-10-13 | 1990-10-13 | Glue membrane for measurement of total of colony |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN90108397A CN1039825C (en) | 1990-10-13 | 1990-10-13 | Glue membrane for measurement of total of colony |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1051758A true CN1051758A (en) | 1991-05-29 |
| CN1039825C CN1039825C (en) | 1998-09-16 |
Family
ID=4880979
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN90108397A Expired - Fee Related CN1039825C (en) | 1990-10-13 | 1990-10-13 | Glue membrane for measurement of total of colony |
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| Country | Link |
|---|---|
| CN (1) | CN1039825C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1063226C (en) * | 1997-12-02 | 2001-03-14 | 中国科学院新疆化学研究所 | Detecting and screening method for biological active additive component of cosmetics |
| CN1908186B (en) * | 2005-08-09 | 2011-05-25 | 沈阳中科靓马生物工程有限公司 | Method of measuring bacteria amount and special agent and apparatus therefor |
| CN105372186A (en) * | 2014-08-18 | 2016-03-02 | 上海复星长征医学科学有限公司 | A method of detecting microorganisms in blood |
| CN106479931A (en) * | 2016-11-04 | 2017-03-08 | 哈尔滨亿隆科技有限公司 | A kind of improved formulations of blood agar culture-medium |
| CN106932383A (en) * | 2015-12-30 | 2017-07-07 | 华南理工大学 | A kind of bacterium total testing pipe and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3903777A1 (en) * | 1989-02-09 | 1990-08-16 | Bruker Analytische Messtechnik | METHOD FOR THE QUICK DETECTION OF MICROORGANISMS IN SAMPLES AND DEVICE FOR IMPLEMENTING THE METHOD |
| FR2649411B1 (en) * | 1989-07-07 | 1996-09-20 | Biosem | QUANTITATIVE DETERMINATION AND IDENTIFICATION OF BACTERIA |
-
1990
- 1990-10-13 CN CN90108397A patent/CN1039825C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1063226C (en) * | 1997-12-02 | 2001-03-14 | 中国科学院新疆化学研究所 | Detecting and screening method for biological active additive component of cosmetics |
| CN1908186B (en) * | 2005-08-09 | 2011-05-25 | 沈阳中科靓马生物工程有限公司 | Method of measuring bacteria amount and special agent and apparatus therefor |
| CN105372186A (en) * | 2014-08-18 | 2016-03-02 | 上海复星长征医学科学有限公司 | A method of detecting microorganisms in blood |
| CN105372186B (en) * | 2014-08-18 | 2018-12-18 | 上海复星长征医学科学有限公司 | The method for detecting microorganism in blood |
| CN106932383A (en) * | 2015-12-30 | 2017-07-07 | 华南理工大学 | A kind of bacterium total testing pipe and preparation method thereof |
| CN106479931A (en) * | 2016-11-04 | 2017-03-08 | 哈尔滨亿隆科技有限公司 | A kind of improved formulations of blood agar culture-medium |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1039825C (en) | 1998-09-16 |
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