Patent ID: 9631243
Date: 2017-04-25
CPC Classifications: C12Q

Claim:
1. A method of detecting the presence or absence of Prototype JC virus in a biological sample from a subject, comprising: amplifying and detecting an amount of a first nucleic acid molecule and an amount of a second nucleic acid molecule from the biological sample using a using a multiplex quantitative real-time polymerase chain reaction (qPCR) assay, comprising: amplifying the first nucleic acid molecule with a first oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of the nucleotide sequences set forth as SEQ ID NOs: 3 (JRR-1) and 4 (JRR-2), respectively; detecting the amplified first nucleic acid molecule with a first oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 5 (JRR-1.1), or the complement thereof; amplifying the second nucleic acid molecule with a second oligonucleotide primer pair comprising forward and reverse oligonucleotide primers consisting essentially of nucleotide sequences set forth as SEQ ID NOs: 6 (JCT-3) and 7 (JCT-4), respectively; detecting the amplified second nucleic acid molecule with a second oligonucleotide probe consisting essentially of the nucleotide sequence set forth as SEQ ID NO: 8 (JCT-1.2), or the complement thereof; and wherein the first nucleic acid molecule is present in an Archetype JC virus genome but not the Prototype JC virus genome, and the second nucleic acid molecule is present in both the Archetype and Prototype JC virus genomes; detecting a greater amount of the second nucleic acid molecule than the first nucleic acid molecule in the biological sample identifies the presence of Prototype JC virus in the biological sample; and detecting an equal amount of the first and second nucleic acid molecules in the biological sample identifies the absence of Prototype JC virus in the biological sample.