Respiratory syncytial virus (rsv) viral load detection assay

Abstract

The invention concerns a method for the extraction of nucleic acids from biological samples e.g. tissue material or sputum derived from human or animal species and the quantitative detection thereafter of said nucleic acids e.g. in terms of viral load, more specifically RSV viral load detection.

1 . A method for the extraction of nucleic acids from a biological sample comprising the following steps:
 a) obtaining said biological sample,   b) adding a phosphate buffered saline comprising about 10% dithiothreitol to said biological sample of step a),   c) incubating under ultrasonic conditions,   d) adding a lysis buffer,   e) obtaining a lysed biological sample, and   f) extracting said nucleic acid from said lysed biological sample.   
     
     
         2 . The method according to  claim 1 , wherein the incubating in step c) is performed with an ultrasonic Covaris S2 apparatus and the extraction in step f) is performed using EasyMAG procedure. 
     
     
         3 . The method according to  claim 1 , wherein said nucleic acid is DNA or RNA from a pathogen. 
     
     
         4 . The method according to  claim 3 , wherein said biological sample is selected from the group consisting of frozen sputum, nasopharyngeal wash (NPW) or lung tissue. 
     
     
         5 . The method according to  claim 1 , further comprising quantifying said nucleic acid. 
     
     
         6 . The method according to  claim 5 , wherein the quantifying said nucleic acid is performed by multi component nucleic acid enzyme technology (MNAzyme). 
     
     
         7 - 8 . (canceled)