Giardia and Cryptosporidium ("two insects") are two pathogenic microorganisms that seriously endanger the safety of drinking water. "Two worms" have the characteristics of small individual, low pathogenicity, simple route and mode of infection, and widespread epidemic distribution. In 2006, China's "Standards for Drinking Water Hygiene" increased the "two insects" test, which stipulated that less than 1 cyst or oocyst per 10L. Today, Xiaobian introduces how to use the special centrifuge for the detection of two insects to carry out the detection of two insects.
First, inoculation and panning and concentration of the sample are carried out, and then centrifugation is started.
a. Centrifuges for the detection of two insects should be placed on a firm workbench or floor to minimize sloshing.
b. Balance the symmetrical centrifuge tube, balance it with the adapter and the centrifugal barrel, and the weight difference should not exceed 0.5g to minimize the swing. Note: If the tube is not balanced, it will generate additional vibration, which will affect the precipitation of the target.
c. Set the centrifuge parameters 2000g, 15min. The sample stops naturally in the centrifuge and cannot be stopped by the brake system.
d. Use a vacuum pump or peristaltic pump to draw the supernatant from the centrifuge tube and install a pipette at the inlet to reduce the vacuum of the inlet. Finally, take 5-8ml (you can mark it on the tube in advance). The flow rate was controlled at 200 ± 20 ml / min. Note that the pipette is placed in the center of the liquid surface, and the supernatant is taken up near the liquid surface to reduce the loss of sedimentation.
e. Mix the remaining concentrate in the centrifuge tube on a vortex mixer for 20 seconds.
f. Rinse the pipette with buffer to transfer all liquid to the L-tube. (Note: The pipette needs to be rinsed with buffer to prevent the pipette wall from hanging due to drying.)
g. Wash the pipette twice with pure water (if the remaining volume is 7ml, 1.5ml each time), and transfer the cleaning solution into the same L-shaped tube. Leave 20 seconds for each rinse and then remove the liquid.
After centrifugation and pipetting of the two-in-one centrifuge , the two insects are captured and stained. Finally, the results of the immunofluorescence test can be used to judge the situation of the two insects.
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