We are trying to study viral strain variants. Our starting material is genomic DNA isolated from oral, blood or nasal rinse. Serological study has given viral positive. We are not able to amplify genes for variant analysis. Of 10 samples only half of the samples give positive pcr amplifications. What could be the problem? Could it be the low copy number of the virus or is it with the pcr protocol?
Is there a given limit of detection for your PCR test?
I worked with a clinical analysis of Helicobacter pyl. in stomach biopsies. The LOD was about 5-10 Hp in the PCR-tube to give a positiv result. I don't know it the tests are compareable, but high content of human (background) DNA can influence the sensitivity and PCR kinetics.
Thank you Gudrun Lang and Marco.... Currently I am dealing with EBV... Seems that the viral load is very low... Is there any means which can improve the qualilty and quantity of the amplicons (even if the copy number is low)... As Gudrun has mention high human DNA content can influence the sensitivity of PCR kinetics.. Do we have any techniques that can masked the human DNA but amplify even the low viral genes...
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