Any Microbiologists out there to help

S

sonar_eit

Registered Member
Hi I am an Engineering PhD student and am looking at detection limits of different assays such as PCR realtime PCR and others. Problem is that I do not know how to equate the following:
1. [nanogram protein antigens per mL] to [spores per mL]
2. [microgram protective antigen per mL] to [spores oer mL]
3. [nanogram antigens per mL] to [spores per mL]
4. and how to find how many fentograms of DNA there are per spore or cell
of a certain bacteria.

I do not know were to find information like this since I am just an Environmental Engineer and have not even seen units like these before. If there is some resource I can find this information could someone help, thank you.

Mark
 
Abstract:

"PCR products were sequenced and the data used to design primers that were directed at the ribosomal RNA genes and internal transcribed spacer regions. Specificity was tested against more than 40 common soil organisms, host plants, and spore suspension contaminants, as well as P. brassicae isolates from around Australia and the world. Sensitivity was determined to be 0.1 fentograms (10–15g) for pure template and as low as 1,000 spores per g of potting mix. In soil, P. brassicae was detected in all soils where the inoculum was sufficient to result in clubroot symptoms. Also outlined is a simple method of DNA extraction from soil." In "Specific Polymerase Chain Reaction Primers for the Detection of Plasmodiophora brassicae in Soil and Water," by R. Faggian, S. R. Bulman, A. C. Lawrie, and I. J. Porter. http://72.14.253.104/search?q=cache...entograms+DNA+spores&hl=en&gl=us&ct=clnk&cd=1

You're converting weight per ML to spores per mL and this would vary between species. There would be no direct way to convert these without knowing the equivalents for each species in question.

10 fentograms of KHV DNA correspond to 30 virions koi herpesvirus (KHV). "From a KHV genomic library, a hypothetical thymidine kinase gene (TK) was identified, subcloned and expressed as a recombinant protein. Preliminary characterization of the recombinant TK showed that it has a kinase activity using dTTP but not dCTP as a substrate. A PCR assay based on primers selected from the defined DNA sequence of the TK gene was developed and resulted in a 409 bp amplified fragment." http://lib.bioinfo.pl/auth:Hedrick,RP

You might trying doing a species-specific search on microsonification for DNA amplification or similar technique. You need a database.
 
Last edited:
Bohm, J., A. Hahn, R. Schubert, G. Bahnweg, N. Adler, J. Nechwatal, R. Oehlmann and W. Osswald. 1999. "Reat-time quantitative PCR: DNA determination in isolated spores of mychorrizal fungas Glomus mossae and monitoring of Phytophthora infestans and Phytophthora citricola in their respective host plant." J. Phytopathol., 147: 409-416.
 
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