Customization: | Available |
---|---|
CAS No.: | 071761 |
Formula: | E. Coli Test Kit |
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Biochemical Identification Kit of Bacteria Instructions
071761-- E. IMViC biochemical identification box (four kinds × 10 times)
I) package of reagents
four kinds of biochemical reaction reagents (Table 1), 10 bottles of 0.85% sterile saline, 1 bottle of 0.5 McFarland turbidity tube, 10 Pcs of disposable pipette.
II) using method of ampoule
Before opening of ampoule, disinfectanting the surface with 75% alcohol cotton, then open the aluminum cover under sterile conditions according to the direction of the arrow .aluminum cover ripped open the vial stopper (Figure 1). All the ampoule can be threw away after use and being autoclaving discarded.
III) the using method of E. coli IMVC biochemical identification box
To further differentiate E. coli IMVC coli from fecal coliform and non-detection results.
inoculated the colony to eosin methylene blue plate from gas-production EC broth tube,culture at 35-37 for 24h, pick suspect colony from the plates and inoculated in nutrient agar slant, culture at 35-37 for 18-24h. Picked bacterial colony from nutrient agar slant and place it into sterile saline, compare with 0.5 McFarland turbidity tube, prepared the bacterial suspension into 0.5McFarland (about 108cfu / mL), Pipet 2 drops (approximately 0.06mL) bacterial suspension and add to each biochemical tube. Put the rubber plug on the ampoule after inoculation, generally it is all-stopper (shown in Figure 2 bottles left), and semi-stoppered (Figure 2 bottles shown right)is necessary for some special situation, standing in the Neto groove (Figure 2), or placed in a suitable bottle rack, and cultured at 35-37 in an incubator, then observe the results show in Table 1. Note: If there are special way of inoculation, incubation time or training, please see the instructions that came with each biochemical identification tube.
Table 1 Biochemical Identification of Escherichia coli and non-Escherichia coli
Product Name |
results to determine |
incubation time(hours) |
Instruction |
|
Positive |
Negative |
|||
Peptone water (tryptophan broth) |
red |
discoloration |
24 |
semi-stoppered ampoule (bottles as shown right side in Figure 2) add 2-3 drops of indole reagent after culture , immediately observe the results.
|
Glucose phosphate peptone water (MR-VP medium) |
MR:red |
discoloration |
96 |
Semi-stoppered ampule (bottles as shown right side in Figure 2)) add 2-3 drops of MR reagent after culture , immediately observe the results.
|
VP: red |
discoloration |
48 |
Semi-stoppered ampule (bottles as shown right side in Figure 2)) add 8 drops of reagent A VP and 4 drops of liquid B liquid after culture, mixing, and then cultured for 10-20 minutes to observe the result
|
|
Koser's citrate |
growth |
no growth |
24-48 |
|
Note: The reagent is placed in the order of HKM catalog from left to right .
IV) biochemical identification procedures E.coli (see below) and biochemical identification features (see Table 2)
Figure E. coli biochemical identification procedures:
Table 2 biochemical characteristics of E. coli
Indole |
MR |
VP |
citrate |
identification (typed) |
+ |
+ |
- |
- |
Typical E.coli |
- |
+ |
- |
- |
Atypical E.coli |
+ |
+ |
- |
+ |
Typical intermediate
|
- |
+ |
- |
+ |
Atypicalintermediate |
- |
- |
+ |
+ |
Typical aerogenes |
+ |
- |
+ |
+ |
Atypical aerogenes |
Note: "+" positive; "-" negative
If there are biochemical reactions occur outside the table, possibly mean impure of sample, should be re-crossed and separated, if necessary, repeat the test.
Packing : 4 types for 10 tests
Details:
1.029030 kovacs indole reagent
2.029040 methyl red reagent
3.029050 Spot Test Veoges-Proskauer reagents( A&B)
A: 60% a- naphthol
B: 40% potassium hydroxide solution
4.075240 peptone water
5.075780 Koser citrate
1.Offering Free Sample for Testing .
2.R & D team to provide technical support.
3.Provide packaging, labeling OEM customization.
Founded in 1993, Guangdong Huankai Microbial Sci. & Tech. Co.,Ltd. is the most important manufacturing and R&D base in the field of detection and control of food safety in China. It is an high-tech corporation belonging to The Guangdong Institute of Microbiology. Through undertaking National and Provincial Level Food safety projects it has won 12 National and Guangdong Provincial level Awards, and has been awarded 33 National patents. In recent years it has been actively participating in the formulation of National and Trading microbial hygiene standards for food and mineral water.
Huankai has always been devoted to monitoring biological safety and controlling the development, production and management of products. At the present it produces microbial monitoring products on a large scale, consisting of 3 large ranges, which amount to nearly 600 varieties of product, including: a series of rapid water quality detection products, highly efficient and environmentally friendly disinfectants as well as biologocial safety laboratory equipment. All of which are largely used in provinces and cities around China, Hong Kong, Macao, Taiwan, as well as other East Asian Countries. After 10 years of development, Huankai has already become the largest manufacturer of microbial monitoring products in China, and the largest manufacturer of industrial cleaning detergent in South China.
With its R&D production bases located in Guangzhou Science City Guangdong Advanced Bio-Tech Demonstration Zone, Huankai now has industry leading GMP workshops producing microbial detection products as well as the largest production line in South China for highly efficient environmentally friendly industrial cleaning detergents used in the food and medical sector. These advanced conditions are helping Huankais speedy development.
Q1:
Sometimes color of dehydrated culture media between batches have subtle differences,would this affect test results?
A1:The source and storage conditions of raw material is different ,so there may be a slight difference in color, which is a normal phenomenon. Products from our company have to undergo a rigorous inspection and sensory biology verification before sale, to ensure that all the indicators characteristics of the product is under enterprise standard to the extent permitted, not affect the test results.
Q2:
After pouring the liquid medium on the plate , medium seems hard to clot or coagulation time is longer, is it some problem with product quality?
A2: The reason may be:
The required hydration in the preparation process have not been done completely .That is to say distilled water is not boil sufficiently to dissolve agar. Because agar proportion, easily settle in the bottom of the bottle, if not fully shaken after high-temperature sterilization, it will lead to uneven upper agar culture gene content and difficult to set.
Q3:
Why some colonies of E. coli chromogenic medium color not shown above, but also confirmed through biochemical tests for E. coli (false negative)?
A3: E. CHROMOGENIC medium is based on the principle of specific enzyme of E. coli and design, 94% of E. coli have β- glucuronidase enzyme, with the chromogenic enzyme substrate medium role in the formation of blue-green colonies. And about 4% of the E. coli does not have β- glucuronidase enzymes, including concern O157: H7 Escherichia coli. Therefore, these E. coli can not be displayed on the color characteristic of E. coli chromogenic medium,it is possible of false-negative on the chromogenic medium. However, the traditional medium of the same can not avoid the problem of false negatives, such as: no gas or slow ferment lactose intolerance may also strain 44.5 false negatives in the conventional medium. For this small part of the E. coli can be detected using other methods.
Q4:
During anaerobic or micro-aerobic culture, is it necessary to remove gas bag from package? How to use oxygen indicator?
A4: During anaerobic or micro-aerobic culture, using method of gassing agent should be done according to the manufacturer's instructions. Remove the paper bag from the plastic packaging, but do not cut the paper bag. Torn the outer packaging of oxygen indicator then it can be used.