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Lee, Shin, Kim, Lee, Kang, Kim, Kim, Song, and Kim: Evaluation of the Real-Q RV Detection Kit for the Identification of Viruses That Result in Respiratory Infections
Original Article

Lab Med Online 2019;9(1):17-21.

Published online: 19 January 2019

DOI: https://doi.org/10.3343/lmo.2019.9.1.17

Evaluation of the Real-Q RV Detection Kit for the Identification of Viruses That Result in Respiratory Infections

Eunyup Lee, M.D., Saeam Shin, M.D., Miyoung Kim, M.D., Young Kyung Lee, M.D., Hee Jung Kang, M.D., Hyun Soo Kim, M.D., Jae-Seok Kim, M.D., Wonkeun Song, M.D., Han-Sung Kim, M.D.

Department of Laboratory Medicine, Hallym University Sacred Heart Hospital, Hallym University College of Medicine, Anyang, Korea

Corresponding author: Han-Sung Kim https://orcid.org/0000-0002-5481-5390 Department of Laboratory Medicine, Hallym University Sacred Heart Hospital, Hallym University College of Medicine, 22 Gwanpyeong-ro 170beon-gil, Dongan-gu, Anyang 14068, Korea Tel: +82-31-380-3932, Fax: +82-31-380-1798, E-mail: kimhs@hallym.or.kr

Received 9 January 2018       Revised 29 January 2018       Accepted 30 January 2018

Copyright © 2019 The Korean Society for Laboratory Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Viral respiratory infections are one of the most common infections worldwide. It is important to detect the virus early and precisely. In this study, we evaluated the limit of detection (LoD) and usefulness of the Real-Q RV Detection kit (BioSewoom, Seoul, Korea). We measured the LoD of the Real-Q RV Detection kit using 10 strains of standard viruses. We then compared the detection results by the Allplex Respiratory Panel Assay kit (Seegene, Seoul, Korea) using 123 clinical specimens. The discrepant results were confirmed by sequencing. Among the 10 standard viruses, the LoD of human rhinovirus (HRV) was the lowest and that of parainfluenza virus 2 and 3 was relatively high as detected by Real-Q RV Detection kit. Agreements of the two kits ranged from 95.9% to 100%. Three specimens detected negative by the Allplex Respiratory Panel kit were detected as adenovirus (AdV) by the Real-Q RV Detection kit and were confirmed by sequencing. Similarly, a specimen detected negative by the Allplex Respiratory Panel kit was detected as HRV by the Real-Q RV Detection kit and was confirmed by sequencing. A specimen detected as human enterovirus by the Allplex Respiratory Panel kit was detected as HRV by the Real-Q RV Detection kit and was confirmed by sequencing. Real-Q RV Detection kit showed good diagnostic performance and can be useful for detecting major viruses that cause respiratory infections.

Keywords: Respiratory virus, Multiplex real-time PCR, Real-Q RV Detection kit

REFERENCES

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Table 1.
The lowest detected concentration of the 10 reference viruses with the Real-Q RV Detection kit
Viruses Pathogenic virus Titers The lowest detected con-
Bank No. (PFU/mL) centration (PFU/mL)
AdV KBPV-VR-1 4.0×108   2.56
CoV OC43 KBPV-VR-8 6.1×105   4.88
CoV 229E KBPV-VR-9 1.0×105   0.16
HRV KBPV-VR-39 1.2×105   0.008
RSV A KBPV-VR-41 2.4×106   0.154
Flu A KBPV-VR-33 8.4×105   6.72
Flu B KBPV-VR-34 1.6×105   0.256
PIV 1 KBPV-VR-44 3.9×105   0.624
PIV 2 KBPV-VR-45 2.0×107 160
PIV 3 KBPV-VR-46 1.6×106 64

Abbreviations: PFU, plaque-forming unit; AdV, Adenovirus; CoV, Human coronavirus; HRV, Human rhinovirus; RSVA, Respiratory syncytial virus A; Flu A, Influenza A virus; Flu B, Influenza B virus; PIV, Parainfluenza virus.

Table 2.
Agreements between the results from the Real-Q RV Detection kit and Allplex Respiratory Panel kit
Viral targets Number of samples Agreement
+/+ -/- +/- -/+ (%)
AdV 0 118 0 5 95.9
CoV NL63 0 122 1 0 99.2
CoV OC43 1 122 0 0 100.0
HRV 27 91 3 2 95.9
RSVA 4 118 1 0 99.2
RSVB 12 111 0 0 100.0
Flu A 1 121 1 0 99.2
PIV 1 5 116 2 0 98.4
PIV 2 7 115 1 0 99.2
PIV 3 1 122 0 0 100.0

+/+, positive for Allplex Respiratory Panel kit (Seegene, Seoul, Korea) and positive for Real-Q RV Detection kit (BioSewoom, Seoul, Korea); -/-, negative for Allplex Respiratory Panel kit and negative for Real-Q RV Detection kit; +/-, positive for Allplex Respiratory Panel kit and negative for Real-Q RV Detection kit; -/+, negative for Allplex Respiratory Panel kit and positive for Real-Q RV Detection kit. Abbreviations: AdV, Adenovirus; CoV, Human coronavirus; HRV, Human rhinovirus; RSVA, Respiratory syncytial virus A; RSVB, Respiratory syncytial virus B; Flu A, Influenza A virus; PIV, Parainfluenza virus.

Table 3.
Sequencing analysis of the discrepant sample results from the Real-Q RV Detection kit and Allplex Respiratory Panel kit
Patients Multiplex real time PCR results Sequencing results Sequence identity (%) Compared sequences from the NCBI database (sequence coverage)
1 Real-Q RV Detection kit∗ AdV AdV 98 MF315029.1 (18890–19097)
  Allplex Respiratory Panel kit† Negative      
2 Real-Q RV Detection kit AdV AdV 99 KY471323.1 (18275–18486)
  Allplex Respiratory Panel kit Negative      
3 Real-Q RV Detection kit AdV AdV 98 MF315029.1 (18890–19096)
  Allplex Respiratory Panel kit Negative      
4 Real-Q RV Detection kit HRV HRV 95 KC306837.1 (215–295)
  Allplex Respiratory Panel kit Negative      
5 Real-Q RV Detection kit HRV HRV 92 KF958308.1 (483–570)
  Allplex Respiratory Panel kit HEV      

Abbreviations: AdV, Adenovirus; HRV, Human rhinovirus; HEV, Human enterovirus.

∗ Real-Q RV Detection kit (BioSewoom, Seoul, Korea);

† Allplex Respiratory Panel kit (Seegene, Seoul, Korea).

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