Korean J Lab Med.  2008 Apr;28(2):109-117. 10.3343/kjlm.2008.28.2.109.

Evaluation of Seeplex(TM) RV Detection Kit for Detecting Rhinovirus, Human Metapneumovirus, and Coronavirus

• Sung H
• Park SJ
• Woo YD
• Choi BH
• Kim MN

• KMID: 854837
• DOI: http://doi.org/10.3343/kjlm.2008.28.2.109

Abstract

BACKGROUND: Direct antigen test (DAT) and culture are primary tests to diagnose infections by respiratory viruses, but are mainly available for the traditional viral pathogens such as respiratory syncytial virus (RSV), influenza virus, parainfluenza virus (PIV), and adenovirus in clinical laboratories. The objective of this study was to evaluate a multiplex reverse transcriptase-PCR method using Seeplex(TM) RV Detection kit (Seegene, Korea) for the detection of rhinovirus, coronavirus, and human metapneumovirus (hMPV). METHODS: From January to May 2007, nasopharyngeal aspirates (NPAs) from pediatric patients negative for culture and DAT of traditional viral pathogens were tested with Seeplex(TM). All the amplicons were directly sequenced and homology of the sequences was searched in the National Center for Biotechnology Information (NCBI) database. Patients' medical records were reviewed for clinical and demographic features. RESULTS: Forty-seven (26.4%) of 178 NPAs were positive: 18 rhinovirus, 15 hMPV, 4 RSV A, 3 coronavirus OC43, 3 influenza virus A, 2 adenovirus, 1 coronavirus NL63, and 1 RSV B. Based on maximum identity, each of the sequences indicating rhinovirus, hMPV, and coronavirus OC43 matched to the corresponding viruses with homology of 94-98%, 96-99%, and 98-100%, respectively. Seeplex(TM) positive patients were 0-11 yr old with a male:female ratio of 1.5:1. Clinical diagnoses included 9 pneumonia, 6 bronchiolitis, 2 cold, 1 asthma exacerbation for rhinovirus; 10 pneumonia, 4 bronchiolitis, and 1 clinical sepsis for hPMV; and 1 pneumonia, 2 croup, and 1 cold for coronavirus. CONCLUSIONS: Multiplex reverse transcriptase-PCR method using Seeplex(TM) RV Detection kit is a reliable test to detect rhinovirus, hMPV, and coronavirus. It may improve the diagnostic sensitivity for RSV, influenza virus, PIV, and adenovirus.

Keyword

Culture; Direct Antigen Test; Multiplex PCR; Respiratory Virus

MeSH Terms

Adolescent
Child
Child, Preschool
Coronavirus/classification/*isolation & purification
Coronavirus 229E, Human/classification/genetics/isolation & purification
Coronavirus OC43, Human/classification/genetics/isolation & purification
Female
Humans
Infant
Infant, Newborn
Male
Metapneumovirus/classification/genetics/*isolation & purification
Phylogeny
Reagent Kits, Diagnostic
Respiratory Tract Infections/*diagnosis/virology
Reverse Transcriptase Polymerase Chain Reaction/*methods
Rhinovirus/classification/genetics/*isolation & purification
Sequence Analysis, DNA

Figure

• 

  Fig. 1. Phylogenetic analysis using Neighbour-Joining method showed a close relationship between the isolates of this study (A. Human rhinovirus [HRV] R1, B. Human metapneumovirus [hMPV] M1, C. Coronavirus [CoV] OC43 C1, D. CoV NL63 C4) and archival strains of corresponding viruses. Distances are presented as a number of substitutions per site (a scale of ‘0.1’ means 0.1 nucleotide substitution per site).

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