Interpretation of Weak-Positive Bands in a Multiplex PCR Using Seeplex RV12 ACE Detection Kit.
- Author:
Yun A JO
1
;
Han Sung KIM
;
Young Kyung LEE
;
Jae Seok KIM
;
Wonkeun SONG
;
Hee Jung KANG
;
Kyu Man LEE
Author Information
1. Department of Laboratory Medicine, Hallym University College of Medicine, Chuncheon, Korea. kimhan@hallym.ac.kr
- Publication Type:Original Article
- Keywords:
Respiratory virus;
Multiplex PCR;
Weak positive
- MeSH:
Adenoviridae;
Biotechnology;
Coronavirus;
Coronavirus OC43, Human;
Electrophoresis;
Humans;
Metapneumovirus;
Multiplex Polymerase Chain Reaction;
Orthomyxoviridae;
Paramyxoviridae Infections;
Polymerase Chain Reaction;
Rhinovirus
- From:Journal of Laboratory Medicine and Quality Assurance
2011;33(2):89-93
- CountryRepublic of Korea
- Language:Korean
-
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.
