Detecting hepatitis A virus and parvovirus PB19 in a blood-derived sample comprises using an amplification assay with probes specific for the viruses and an internal control

摘要

Detecting hepatitis A virus (HAV) and parvovirus PB19 in a blood-derived sample comprises extracting viral nucleic acids using a lysis buffer containing an internal control nucleic acid; adding the extract to a PCR mastermix comprising an HAV-specific set of primers and probes, a PB19-specific set of primers and probes and a probe specific for the internal control, where each probe is labeled with two fluorescent dyes; performing nucleic acid amplification; and detecting amplified viral nucleic acids and internal control nucleic acid. Detecting hepatitis A virus (HAV) and parvovirus PB19 in a blood-derived sample comprises concentrating the sample; extracting viral nucleic acids using a lysis buffer containing an internal control nucleic acid and using heated nuclease-free water for elution; adding the extract to a PCR mastermix comprising an HAV-specific set of oligonucleotides (sense and antisense primers, first and second probes), a PB19-specific set of oligonucleotides (sense and antisense primers, third and fourth probes) and a fifth probe specific for the internal control, where each probe is labeled with two fluorescent dyes; performing nucleic acid amplification; and detecting amplified viral nucleic acids and internal control nucleic acid. Independent claims are also included for: (1) composition for detecting hepatitis A virus, comprising oligonucleotides with SEQ ID NO:3-6 (defined sequences given in the specification); (2) composition for detecting parvovirus PB19, comprising oligonucleotides with SEQ ID NO:8-11 (defined sequences given in the specification); (3) composition for detecting hepatitis A virus and parvovirus PB19, comprising oligonucleotides with SEQ ID NO:3-6 and oligonucleotides with SEQ ID NO:8-11.