Evaluation of a novel real-time RT-PCR using TOCE technology compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses

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Abstract

Background: Various kinds of commercial molecular systems have been developed for fast and more accurate detection of respiratory viruses. Anyplex™ II RV16 [RV16] was designed for simultaneous detection of 16 respiratory viruses using multiplex PCR coupled with TOCE™ technology. Objectives: To compare the performance of RV16 with those of culture and Seeplex® RV15 ACE [RV15] by determining their sensitivity and specificity. Study design: Seven hundred and thirty respiratory samples were tested by modified shell vial culture method, RV16, and RV15. For molecular tests, automated nucleic acid extraction and liquid handling system using MICROLAB Nimbus IVD (Hamilton, USA) was adopted to maximize the workflow and accuracy. Performance of each assay was determined against a composite reference standard. Results: Two hundred and one samples (28%) out of 730 samples were positive by culture, while additional 281 (39%) were positive by RV16 or RV15. Sensitivities of RV16, RV15, and culture for virus tested were as follows: 100/93/63% for influenza A, 90/80/69% for influenza B, 98/94/63% for RSV, 98/52/23% for adenovirus, and 100/75/46% for PIV. For viruses not covered by culture, sensitivities of RV16 and RV15 were as follows: 99/81% for rhinovirus, 92/100% for coronavirus OC43, 100/56% for coronavirus 229E/NL63, 92/88% for metapneumovirus, 100/62% for bocavirus, and 91/91% for enterovirus. Overall, the specificities of culture, RV16, and RV15 (Seegene) were 100/99.9/99.9%. Conclusions: RV16 assay was superior to culture method and RV15 and will be a promising tool for patient management and public health epidemiology.

Original languageEnglish
Pages (from-to)338-342
Number of pages5
JournalJournal of Clinical Virology
Volume57
Issue number4
DOIs
Publication statusPublished - 2013 Aug 1

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Real-Time Polymerase Chain Reaction
Viruses
Technology
Coronavirus
Human Influenza
Bocavirus
Metapneumovirus
Rhinovirus
Workflow
Enterovirus
Multiplex Polymerase Chain Reaction
Adenoviridae
Nucleic Acids
Epidemiology
Public Health
Sensitivity and Specificity

Keywords

  • Multiplex RT PCR
  • Real-time
  • Respiratory virus
  • Viral isolation

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

@article{bfdcef49512443e2a74694dd427e2d01,
title = "Evaluation of a novel real-time RT-PCR using TOCE technology compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses",
abstract = "Background: Various kinds of commercial molecular systems have been developed for fast and more accurate detection of respiratory viruses. Anyplex™ II RV16 [RV16] was designed for simultaneous detection of 16 respiratory viruses using multiplex PCR coupled with TOCE™ technology. Objectives: To compare the performance of RV16 with those of culture and Seeplex{\circledR} RV15 ACE [RV15] by determining their sensitivity and specificity. Study design: Seven hundred and thirty respiratory samples were tested by modified shell vial culture method, RV16, and RV15. For molecular tests, automated nucleic acid extraction and liquid handling system using MICROLAB Nimbus IVD (Hamilton, USA) was adopted to maximize the workflow and accuracy. Performance of each assay was determined against a composite reference standard. Results: Two hundred and one samples (28{\%}) out of 730 samples were positive by culture, while additional 281 (39{\%}) were positive by RV16 or RV15. Sensitivities of RV16, RV15, and culture for virus tested were as follows: 100/93/63{\%} for influenza A, 90/80/69{\%} for influenza B, 98/94/63{\%} for RSV, 98/52/23{\%} for adenovirus, and 100/75/46{\%} for PIV. For viruses not covered by culture, sensitivities of RV16 and RV15 were as follows: 99/81{\%} for rhinovirus, 92/100{\%} for coronavirus OC43, 100/56{\%} for coronavirus 229E/NL63, 92/88{\%} for metapneumovirus, 100/62{\%} for bocavirus, and 91/91{\%} for enterovirus. Overall, the specificities of culture, RV16, and RV15 (Seegene) were 100/99.9/99.9{\%}. Conclusions: RV16 assay was superior to culture method and RV15 and will be a promising tool for patient management and public health epidemiology.",
keywords = "Multiplex RT PCR, Real-time, Respiratory virus, Viral isolation",
author = "Cho, {Chy Hyun} and Bayarjavkhlan Chulten and Lee, {Chang Kyu} and Myung-Hyun Nam and Soo-Young Yoon and Lim, {Chae Seung} and Yunjung Cho and Kim, {Young Kee}",
year = "2013",
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doi = "10.1016/j.jcv.2013.04.014",
language = "English",
volume = "57",
pages = "338--342",
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T1 - Evaluation of a novel real-time RT-PCR using TOCE technology compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses

AU - Cho, Chy Hyun

AU - Chulten, Bayarjavkhlan

AU - Lee, Chang Kyu

AU - Nam, Myung-Hyun

AU - Yoon, Soo-Young

AU - Lim, Chae Seung

AU - Cho, Yunjung

AU - Kim, Young Kee

PY - 2013/8/1

Y1 - 2013/8/1

N2 - Background: Various kinds of commercial molecular systems have been developed for fast and more accurate detection of respiratory viruses. Anyplex™ II RV16 [RV16] was designed for simultaneous detection of 16 respiratory viruses using multiplex PCR coupled with TOCE™ technology. Objectives: To compare the performance of RV16 with those of culture and Seeplex® RV15 ACE [RV15] by determining their sensitivity and specificity. Study design: Seven hundred and thirty respiratory samples were tested by modified shell vial culture method, RV16, and RV15. For molecular tests, automated nucleic acid extraction and liquid handling system using MICROLAB Nimbus IVD (Hamilton, USA) was adopted to maximize the workflow and accuracy. Performance of each assay was determined against a composite reference standard. Results: Two hundred and one samples (28%) out of 730 samples were positive by culture, while additional 281 (39%) were positive by RV16 or RV15. Sensitivities of RV16, RV15, and culture for virus tested were as follows: 100/93/63% for influenza A, 90/80/69% for influenza B, 98/94/63% for RSV, 98/52/23% for adenovirus, and 100/75/46% for PIV. For viruses not covered by culture, sensitivities of RV16 and RV15 were as follows: 99/81% for rhinovirus, 92/100% for coronavirus OC43, 100/56% for coronavirus 229E/NL63, 92/88% for metapneumovirus, 100/62% for bocavirus, and 91/91% for enterovirus. Overall, the specificities of culture, RV16, and RV15 (Seegene) were 100/99.9/99.9%. Conclusions: RV16 assay was superior to culture method and RV15 and will be a promising tool for patient management and public health epidemiology.

AB - Background: Various kinds of commercial molecular systems have been developed for fast and more accurate detection of respiratory viruses. Anyplex™ II RV16 [RV16] was designed for simultaneous detection of 16 respiratory viruses using multiplex PCR coupled with TOCE™ technology. Objectives: To compare the performance of RV16 with those of culture and Seeplex® RV15 ACE [RV15] by determining their sensitivity and specificity. Study design: Seven hundred and thirty respiratory samples were tested by modified shell vial culture method, RV16, and RV15. For molecular tests, automated nucleic acid extraction and liquid handling system using MICROLAB Nimbus IVD (Hamilton, USA) was adopted to maximize the workflow and accuracy. Performance of each assay was determined against a composite reference standard. Results: Two hundred and one samples (28%) out of 730 samples were positive by culture, while additional 281 (39%) were positive by RV16 or RV15. Sensitivities of RV16, RV15, and culture for virus tested were as follows: 100/93/63% for influenza A, 90/80/69% for influenza B, 98/94/63% for RSV, 98/52/23% for adenovirus, and 100/75/46% for PIV. For viruses not covered by culture, sensitivities of RV16 and RV15 were as follows: 99/81% for rhinovirus, 92/100% for coronavirus OC43, 100/56% for coronavirus 229E/NL63, 92/88% for metapneumovirus, 100/62% for bocavirus, and 91/91% for enterovirus. Overall, the specificities of culture, RV16, and RV15 (Seegene) were 100/99.9/99.9%. Conclusions: RV16 assay was superior to culture method and RV15 and will be a promising tool for patient management and public health epidemiology.

KW - Multiplex RT PCR

KW - Real-time

KW - Respiratory virus

KW - Viral isolation

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