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QUANTITATIVE DETERMINATION

 

Quantitative determination of the stool´s infestation with parasites

Sedimentation method for human and veterinary medicine

 

Procedure I (open weighing)

Unscrew the cap of Processing tube I. Put the cap with the plunger on a balance with the plunger placed upwards, the cap downwards.
1 2 3
Weigh the plunger of tube I before the sample is taken. Weigh the plunger of tube I after the sample is taken.

The difference in weigh corresponds to the weigh of the sample: 0,80 g.

 
Full processing as described in user manual by ParasiTrap®.
4 5
Processing tube I loaded with sample

 
0,5-1,0 ml
6 7
Concentrated parasites on the bottom

 

 Dilution of parasites up to 1 ml volume

 
8 9

Take out 100 µl and count the parasites in microscope slides or in counting chamber
.

 

Procedure II (closed weighing)
1 2 3
Tube I with Medium A is weighed without sample IMPORTANT
while loading the tube with sample, no medium should get lost! 		Tube I with Medium A and the samle is weighed again
The difference in weigh corresponds to the weigh of the sample: 0,70 g.

 
Full processing as described in user manual by ParasiTrap®.
4 5
Processing tube I loaded with sample

 
0,5-1,0 ml
6 7
Concentrated parasites on the bottom

 

 Dilution of parasites up to 1 ml volume

 
8 9

Take out 100 µl and count the parasites in microscope slides or in counting chamber

Evaulation for the ParasiTrap sedimentation method:

The parasite concentrate/sediment obtained after completion of this procedure is diluted respectively adjusted to 0,5 ml or 1,0 ml. Mix thoroughly and then aspirate 100 µl by a tuberculin syringe or micro pipette and count the number of parasites contained in it. The number of parasites - multipled by 5 (for 0,5 ml of sediment) or 10 (for 1,0 ml of sediment) - and the known amount of sample allow to determine the parasite concentration per gram of stool easily.

CAUTION: it is recommended to have several samples. The values are only correct, if the sample has been suspended respectively processed with extreme care.

 

Flotation method for human and veterinary medicine

 

Procedure I (open weighing)

Unscrew the cap of Processing tube I. Put the cap with the plunger on a balance with the plunger placed upwards, the cap downwards.

 
1 2 3
Weigh the plunger of tube I before the sample is taken Weigh the plunger of tube I after the sample is taken

The difference in weigh corresponds to the weigh of the sample: 0,80 g.

 

Full processing as described in user manual by ParasiTrap® flotation.
4 5 6
Processing tube I loaded with sample

 

 Concentrated parasites on the meniscus surface

 
7 8

Collect all parasites from the upper layer of meniscus and count they on microscope slides or in counting chamber.

.

 

Procedure II (closed weighing)

1 2 3
Tube I with Medium A is weighed without sample IMPORTANT
while loading the tube with sample, no medium should get lost! 		Tube I with Medium A and the sample weighed again

The difference in weigh corresponds to the weigh of the sample: 0,70 g.

 

Full processing as described in user manual by ParasiTrap® flotation.
4 5 6
Processing tube I loaded with sample

 

 Concentrated parasites on the meniscus surface

 
7 8
Collect all parasites from the upper layer of meniscus and count they on microscope slides or in counting chamber.

The known amount of sample allow to determine the parasite concentration per gram of stool easily.

CAUTION: it is recommended to have several samples. The values are only correct, if the sample has been suspended respectively processed with extreme care.

The better the faecal suspension the better the results.
Condition for a successful examination is that the sample has to be fresh as possible.
Enhance your chance for successful examination:
Take samples from different parts of faecal material
Examine sample from different days
Two examinations are better than one
Samples processed by two different diagnostic methods - flotation and sedimentation - increasing qualitative and quantitative the number of parasites.
Use cover glass size 24x60 mm. Examine the cover-slip on the slide systematically and thoroughly.
Liquid faecal samples:
Use Pasteur pipette to take 1 or 2 ml sample. Use the closed weighing method.

For further information see to corresponding literature about parasitology.

 

 

Sedimentation method for veterinary medicine

BIG ANIMALS

Procedure

1 2 3
Weigh the sample vessel before the faecal material is taken (with cap) Insert 2x10 cm² sample into the sample vessel and intensively stirring with the wooden spoon Weigh the vessel after the sample is taken (with cap)
The difference in weigh corresponds to the weigh of the sample (13,0 g)

 
4 5
First filtration of faecal suspension

 

 Determination of the filtrate´s volume. Adjust the filtrate to 30, 35 or maximal 40 ml.

 
6 7
Shake intensively

 

 Fill to the processing tube I 2 ml suspension without a waiting period

 
8 9 10
Add 1,5 ml Combi Medium for separation and staining Screw the processing tube II onto the tube I. The assembled system is rotated 180º Mixed by shaker at max. rotary speed for approx. 10-15 sec. 
If you have not shaker, shake it by hand for 30 sec.

 
11 12
After that the sample in the upper vessel is transferred into the lower tube II by shaking in vertical direction

 

 The filtered suspension is centrifuged for 5 min. at 1000-1500 g (separation of debris)

 
13 14
Remove the tube I with the filter-piece

 

 The top layer which is solid has to be detached from tube´s wall by a cotton wool stick

 
15 16
Concentrated parasites are in the bottom

 

 Fill the concentrated parasites up to 0,5 or 1,0 ml with Medium A

 
17 18
Take out immediately 100 µl material after the resuspension... ...and count the parasites. If necessary, use for counting 2 or 3 slides. 
Make sure that your preparation is not too thick. You must be able to read through your preparation clearly. Don´t let dry your slides before examination.

 
19
Microscopical examination

 

Flotation method for veterinary medicine

BIG ANIMALS

Procedure
1 2 3
Weigh the sample vessel before the faecal material is taken (with cap) Insert 2x10 cm² sample into the sample vessel and intensively stirring with the wooden spoon Weigh the vessel after the sample is taken (with cap)
The difference in weigh corresponds to the weigh of the sample (13,0 g)

 
4 5
First filtration of faecal suspension

 

 Determination of the filtrate´s volume. Adjust the total volume with Flotation PLUS solution between 30, 35 or maximal 40 ml.

 
6 7
Shake intensively

 

 Fill to the processing tube I 5 ml suspension without a waiting period

 
8 9 10
Screw the processing tube II onto the tube I. The assembled system is rotated 180º The filtered suspension is centrifuged for 5 min. at 1000-1500 g (separation of debris) After that the sample in the upper vessel is transferred into the lower tube II by shaking in vertical direction 
If you have not shaker, shake it by hand for 30 sec.

 
11 12
The filtered suspension is centrifuged for 5 min. at 250-300 g

 

 Tube II filled with ParasiTrap® Flotation PLUS solution to form a slight positive meniscus

 
13
Wait 20-30 min. to concentrate parasites in the upper layer of meniscus (flotation step)

Alternative methods sampling the reverse meniscus added cover-slip or glass slide, take a loop, glass rod or fine pipette. Count the parasites on the glass slide or in a chamber.

Liquid faecal samples:
Take 10, 15 or maximum 20 ml sample with a pipette.

 

 

 

 

 

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