Patent ID: 7985541
Filing Date: 2011-07-26
Classification: C12N,C12Q

Abstract:
1. A method for quantitating newly initiated RNA of a positive strand RNA virus comprising: contacting an isolated replicase complex for the positive strand RNA virus, an isolated viral replicon template RNA for the positive strand RNA virus, nucleotides, and a labeled nucleotide analog, and 2′-O-methyl-5-methyluridine-5′- triphosphate under conditions sufficient for in vitro RNA synthesis, to form a newly synthesized RNA population comprising the labeled nucleotide analog; hybridizing a probe and the newly synthesized RNA population comprising the labeled nucleotide analog, under stringent hybridization conditions, wherein the probe is complementary to at least a portion of a transcription initiation region of the newly synthesized RNA population; digesting unhybridized, single-stranded RNA with a single-strand specific ribonuclease to form a protected RNA population comprising the labeled nucleotide analog; detecting the protected RNA population comprising the labeled nucleotide analog; and quantitating the protected RNA population comprising the labeled nucleotide analog.