The project was able to show that use of the conventional PCR approach using DNA as a target showed positive amplification detection of non-viable cells of Vibrio cholerae which, in many instances, would be considered a false-positive result.  Total RNA from both viable and non-viable cells were purified and treated with DNAse (RNAse-free) to avoid any amplification from the contaminating target DNA. A RT-PCR approach using this rapid and effective method for RNA purification showed amplification of the target RNA only from the viable cells. The sensitivity of detection of viable cells of V. cholerae was greater than or equal to 103, which is well within the minimum number of cells required for infection. The use of a reliable prokaryotic RNA extraction method followed by RT-PCR amplification of the target RNA can be used for specific detection of viable microbial pathogens such as V. cholerae, avoiding undesired false-positive results.