Sample preparation for difficult sample types

Abstract

Devices and methods are provided for collecting and handling difficult sample types.

1 . A method of amplifying RNA from a direct blood sample comprising:
 (a) adding the direct blood sample to a sample buffer,   (b) bead beating the sample buffer,   (c) extracting the RNA from the sample buffer, and   (d) amplifying the RNA.   
     
     
         2 . The method of  claim 1 , wherein steps (b) through (d) are performed in a closed sample vessel. 
     
     
         3 . The method of  claim 1 , wherein steps (a) through (d) are performed without centrifugation, ethanol precipitation, and DNA digestion. 
     
     
         4 . The method of  claim 1 , wherein the RNA is host RNA. 
     
     
         5 . The method of  claim 1 , wherein the direct blood sample is collected in a RNA stabilized solution. 
     
     
         6 . The method of  claim 1 , wherein the direct blood sample is collected in a non-RNA stabilizing solution. 
     
     
         7 . The method of  claim 1 , wherein the direct blood sample is separated prior to step (a) and a blood fraction is used. 
     
     
         8 . The method of  claim 1 , further comprising filtering the sample between steps (a) and (b). 
     
     
         9 . The method of  claim 1 , wherein the RNA is provided in high copy number. 
     
     
         10 . The method of  claim 1 , wherein the RNA is from microorganisms. 
     
     
         11 . The method of  claim 1 , further comprising washing the RNA following the extracting step, wherein the washing step comprises no more than three washes. 
     
     
         12 . The method of  claim 1 , wherein the sample buffer includes a guanidinium compound. 
     
     
         13 . The method of  claim 1 , wherein step (g) includes a first-stage multiplex PCR and a plurality of individual second-stage PCR reactions.