Some of my colleagues infected ducks with H5N1 virus, but they collected organs from one animal per time point. This means that we do not have biological replicate.
How can I measure relatively or absolutely the virus titer in these organs without having biological replicates ?
Hi Mohamed, you need more than 1 animal. I think the experiment must be repeated. I was working with Kathy Magor(she was doing Infections of ducks with influenza) and she always used three animals for one time point. For measuring the viral titer you could made qRT-PCR of the influenza matrix gene.
Unfortunately, the short answer is that you cannot undertake a statistical analysis if n=1. Like with most assays, technical replicates do substitute for biological replicates. To get a meaningful result you must collect samples from several animals for each time point. In addition, I find that virus infection in vivo is highly variable. When using in bred mice, I find that it is necessary to sample at least 6 samples per time point, and up to 10 samples per timepoint in some cases. This is because virus infection progresses at very different rates in different animals, despite using the same dose for inoculation. If you are using out-bred ducks, you should expect even more variability.
I have just re- read my response and released that I missed out an important word; "Not". (!!!)
It should have read:
"Like with most assays, technical replicates do NOT substitute for biological replicates. "
:-)
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