COMPOSITIONS AND METHODS FOR USE IN A PCR ASSAY FOR DETERMINING THE GENOTYPE AND VIRAL LOAD FOR RESPIRATORY SYNCYTIAL VIRUS

摘要

Methods are provided for a sensitive and specific assay for the determination of viral load and genotyping of RSV in a biological sample. Compositions and kits for use in the methods also are provided, including optimized primers for the amplification of and detection of the RSV open reading frames from subtypes A and B, and probes for distinguishing between the subtypes. Also provided are methods for amplifying and sequencing an open reading from of an RSV F protein, and compositions and kits for use in the methods.

主权项

1. A method for simultaneously determining the viral load and genotype of a respiratory syncytial virus (RSV) in a biological sample, comprising:
a) amplifying a nucleic acid sequence encoding an RSV open reading frame (ORF) in a biological sample using a set of primers that specifically amplifies an RSV A ORF and/or an RSV B ORF and at least two nucleic acid probes in a quantitative polymerase chain reaction assay, wherein at least one probe specifically binds to an RSV subtype A nucleic acid sequence and at least one probe specifically binds to an RSV subtype B nucleic acid sequence, and wherein the at least two nucleic acid probes are differentially labeled; b) determining the viral load and the RSV subtype(s) present in the biological sample.