Can anyone tell me how long it is good to store animal blood without reducing the parasite detecting efficiency? I tried isolating whole blood DNA after 2 months of storage and it is not giving any results as compared to DNA isolated after 2 days. If there is a different method to store for parasitic study please tell that too.
Use EDTA vacutainer when collecting the blood, freezing it at -20 C will be more than enough to keep the DNA intact.
Thanks Raditijo.. I am already using edta vacutainer but not storing blood at -20 C. I have kept it at 4C due to avoid frequent freeze thaw.. but as u suggest i will try to store at -20 C..
Hello, in our lab we store the blood sample in buffer SET (1 M Tris- HCl pH 8.0, 0.5 M EDTA, 10% SDS), in field we have the sample to 15-20 °C for 20 days, but in the lab are stored to -20°C. The success of the amplification depends of the intensity of infection, we are incapable to amplify hemosporidian parasites if we found a only one parasite in one smear. If the complete smear have 1-4 parasites, we extract DNA with comercial kits. If the number of parasites are greater, the extraction with phenol-cloroform is exellent, cheep and successful.
Thanks Angie,
I have spotted 10-15 parasite per filed view. for an average of 40-50 infected WBCs for my parasite. In spite of such high infection intensity, the detection value of PCR degrades with time at 4 C not allowing me to store my sample beyond two months. But i will look into your suggestion and will try to store my sample in SET buffer next time to check if it works.
Thanks again.
IN ALL DETECTION TECHNIQUE OF BLOOD PARASITE YOU MUST AVOID FREEZING AND THAWING WHICH MIGHT BE LEAD TO PARASITE DESTRUCTION
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