Interpretation of Weak-Positive Bands in a Multiplex PCR Using Seeplex RV12 ACE Detection Kit / 임상검사와정도관리

ABSTRACT

BACKGROUND: Multiplex PCR assay is a sensitive tool for the detection of various respiratory viruses. Seeplex RV12 ACE Detection (Seegene, Korea) assay is a multiplex RT-PCR assay for detection of 12 respiratory viruses. We had observed several cases with faint bands in the test. Those results were investigated in this study. METHODS: A total of 163 specimens were tested with Seeplex RV12 ACE Detection assay. The amplicons showing faint band in electrophoresis were reamplified and sequenced. RESULTS: A total of 99 viruses were detected in 80 specimens (49.1%). Twenty-four amplicons showed faint band in eletrophoresis. All of influenza virus A, parainfluenza viruses (PIV), coronavirus OC43, human metapneumovirus (HMPV) and adenovirus amplicons were reamplified, but 4 of 12 human rhinovirus amplicons were not reamplified. Sequences of reamplified PCR products showed homology of 95-99% to those of corresponding viruses in the National Center for Biotechnology Information database. CONCLUSIONS: Faint bands of influenza virus A, PIV-1, PIV-3, coronavirus OC43, HMPV and adenovirus in Seeplex RV12 ACE Detection assay are specific bands and seems to be weak positive results.