Hello, could someone enlighten me? i have a question regarding the sensitivity of our primer can detect, for example, if the presence of the pathogen is only 1 cell/ 50 mg then we perform DNA extraction. are there any methods to check the sensitivity and specificity of our primer? thank you so much
primer design Ensure the primers are specific to the pathogen’s DNA sequence.
Hanifa Rizky The specificity and sensitivity of PCR analysis are routine tests for any validated PCR kit or primer pair. For specificity, PCR primers are tested against three panels of organisms: target organisms, phylogenetically related organisms, and unrelated others that often occur in natural samples. Sensitivity is tested using a panel of serial dilutions of target DNA or cells, with concentrations validated using other methods. The detection of one cell per 50 mg of material is a low probability event for any detection method.
Hello 1 cell is 1 Copy of DNA. The PCR detection of 1 copy of DNA is not reliable.
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