Journal List > Ann Clin Microbiol > v.22(2) > 1127921

Uwizeyimana, Kim, Kim, Byun, and Yong: Comparison of Multiplex Real-Time Polymerase Chain Reaction Assays for Detection of Respiratory Viruses in Nasopharyngeal Specimens

Abstract

Background

Respiratory tract infections are major public health threats, and the identification of their causative microbes helps clinicians to initiate timely and appropriate antimicrobial therapy and prevent the secondary spread of infection. The main goal of this study was to compare two multiplex real-time polymerase chain reaction (PCR) assays used to detect respiratory viral pathogens in nasopharyngeal swab specimens.

Methods

Between September and October 2017, a total of 84 nasopharyngeal specimens were obtained consecutively from patients in a tertiary hospital using a flocked swab with 3 mL universal transport medium (COPAN Diagnostics, USA). A total of 64 positive and 20 negative sample results from the LG AdvanSure RV real-time RT-PCR kit (LG Life Sciences, Korea) were further retested using a new AdvanSure RV-plus a real-time RT-PCR kit to compare their performance.

Results

Statistical analysis of positive and negative agreement between the two different kits was conducted between the newly introduced AdvanSure RV-plus real-time RT-PCR kit and the AdvanSure RV real-time RT-PCR. The overall agreement was 96.4%, with positive agreement of 98.4% and negative agreement of 90%. The evaluated sensitivity and specificity of AdvanSure RV-plus real-time RT-PCR were 96.9% and 94.7%, respectively, with a kappa value of 0.9 (P<0.001).

Conclusion

The performances of LG AdvanSure RV real-time RT-PCR and the new AdvanSure RV-plus real-time RT-PCR kit showed strong overall agreement. AdvanSure RV-plus real-time RT-PCR had a better detection rate and could detect coronavirus 229E and enterovirus, especially with a high detection rate in coinfection. AdvanSure RV-plus real-time RT-PCR can be considered a useful tool for respiratory virus diagnosis in clinical laboratories.

References

1. Moesker FM, van Kampen JJ, van Rossum AM, de Hoog M, Koopmans MP, Osterhaus AD, et al. Viruses as sole causative agents of severe acute respiratory tract infections in children. PLoS One. 2016; 11:e0150776.
crossref
2. Tregoning JS, Schwarze J. Respiratory viral infections in infants: causes, clinical symptoms, virology, and immunology. Clin Microbiol Rev. 2010; 23:74–98.
crossref
3. Legand A, Briand S, Shindo N, Brooks WA, de Jong MD, Farrar J, et al. Addressing the public health burden of respiratory viruses: the Battle against Respiratory Viruses (BRaVe) initiative. Future Virol. 2013; 8:953–68.
crossref
4. Pinky L and Dobrovolny HM. Coinfections of the respiratory tract: viral competition for resources. PLoS One. 2016; 11:e0155589.
crossref
5. Mehta S, Shin H, Burnett R, North T, Cohen AJ. Ambient particulate air pollution and acute lower respiratory infections: a systematic review and implications for estimating the global burden of disease. Air Qual Atmos Health. 2013; 6:69–83.
crossref
6. Salez N, Vabret A, Leruez-Ville M, Andreoletti L, Carrat F, Renois F, et al. Evaluation of four commercial multiplex molecular tests for the diagnosis of acute respiratory infections. PLoS One. 2015; 10:e0130378.
crossref
7. Peaper DR and Landry ML. Rapid diagnosis of influenza: state of the art. Clin Lab Med. 2014; 34:365–85.
8. Hodinka RL and Kaiser L. Point-counterpoint: is the era of viral culture over in the clinical microbiology laboratory? J Clin Microbiol. 2013; 51:2–8.
9. Basile K, Kok J, Dwyer DE. Point-of-care diagnostics for respiratory viral infections. Expert Rev Mol Diagn. 2018; 18:75–83.
crossref
10. Souf S. Recent advances in diagnostic testing for viral infections. Biosci Horiz. 2016; 9:hwz010.
11. Loeffelholz MJ. Rapid diagnosis of viral infections. Lab Med. 2002; 33:283–6.
crossref
12. Gowin E, Bartkowska-Ś niatkowska A, Joń czyk-Potoczna K, Wysocka-Leszczyń ska J, Bobkowski W, Fichna P, et al. Assessment of the usefulness of multiplex realtime PCR tests in the diagnostic and therapeutic process of pneumonia in hospitalized children: a singlecenter experience. Biomed Res Int. 2017; 2017:8037963.
crossref
13. Huang HS, Tsai CL, Chang J, Hsu TC, Lin S, Lee CC. Multiplex PCR system for the rapid diagnosis of respiratory virus infection: systematic review and metaanalysis. Clin Microbiol Infect. 2018; 24:1055–63.
crossref
14. Kralik P and Ricchi M. A basic guide to real time PCR in microbial diagnostics: definitions, parameters, and everything. Front Microbiol. 2017; 8:108.
crossref
15. Krause JC, Panning M, Hengel H, Henneke P. The role of multiplex PCR in respiratory tract infections in children. Dtsch Arztebl Int. 2014; 111:639–45.
crossref
16. To KK, Lu L, Yip CC, Poon RW, Fung AM, Cheng A, et al. Additional molecular testing of saliva specimens improves the detection of respiratory viruses. Emerg Microbes Infect. 2017; 6:e49.
crossref
17. Cho CH, Lee CK, Nam MH, Yoon SY, Lim CS, Cho Y, et al. Evaluation of the AdvanSure TM realtime RT-PCR compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses. Diagn Microbiol Infect Dis. 2014; 79:14–8.
18. Kuan CS, Yew SM, Hooi PS, Lee LM, Ng KP. Detection of respiratory viruses from ARTI patients by xTAG RVP Fast v2 assay and conventional methods. Malays J Med Sci. 2017; 24:33–43.
19. Piltcher OB, Kosugi EM, Sakano E, Mion O, Testa JRG, Romano FR, et al. How to avoid the inappropriate use of antibiotics in upper respiratory tract infections? A position statement from an expert panel. Braz J Otorhinolaryngol. 2018; 84:265–79.
crossref
20. Marshall DJ, Reisdorf E, Harms G, Beaty E, Moser MJ, Lee WM, et al. Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting. J Clin Microbiol. 2007; 45:3875–82.
crossref
21. Kim H, Hur M, Moon HW, Yun YM, Cho HC. Comparison of two multiplex PCR assays for the detection of respiratory viral infections. Clin Respir J. 2014; 8:391–6.
crossref
22. Bibby DF, McElarney I, Breuer J, Clark DA. Comparative evaluation of the Seegene Seeplex RV15 and realtime PCR for respiratory virus detection. J Med Virol. 2011; 83:1469–75.
crossref
23. Christaki E. New technologies in predicting, preventing and controlling emerging infectious diseases. Virulence. 2015; 6:558–65.
crossref
24. Jung YJ, Kwon HJ, Huh HJ, Ki CS, Lee NY, Kim JW. Comparison of the AdvanSure TM realtime RT-PCR and Seeplex® RV12 ACE assay for the detection of respiratory viruses. J Virol Methods. 2015; 224:42–6.
25. Boivin G, Côté S, Déry P, De Serres G, Bergeron MG. Multiplex realtime PCR assay for detection of influenza and human respiratory syncytial viruses. J Clin Microbiol. 2004; 42:45–51.
crossref
26. Yun SG, Kim MY, Choi JM, Lee CK, Lim CS, Cho Y, et al. Comparison of three multiplex PCR assays for detection of respiratory viruses: Anyplex II RV16, AdvanSure RV, and Real-Q RV. J Clin Lab Anal 2018. doi: 10.1002/jcla.22230.
27. Huh HJ, Kim JY, Kwon HJ, Yun SA, Lee MK, Lee NY, et al. Performance evaluation of Allplex respiratory panels 1, 2, and 3 for detection of respiratory viruses and influenza A virus subtypes. J Clin Microbiol. 2017; 55:479–84.
crossref
28. Yan L, Meng B, Xiang J, Wilson IA, Yang B. Crystal structure of the post-fusion core of the Human coronavirus 229E spike protein at 1.86 Å resolution. Acta Crystallogr D Struct Biol. 2018; 74:841–51.
29. Asner SA, Petrich A, Hamid JS, Mertz D, Richardson SE, Smieja M. Clinical severity of rhinovirus/enterovirus compared to other respiratory viruses in children. Influenza Other Respir Viruses. 2014; 8:436–42.
crossref

Table 1.
Overall sensitivity comparison table
Target virus No. (%) of positives
Total true positive AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR Statistical difference
Adeno 11 11 (100) 10 (90.9) NS
229E 2 2 (100)
BoV 11 11 (100) 10 (90.9) NS
ENT 5 5 (100)
INFA 7 7 (100) 7 (100) NS
MPV 9 9 (100) 8 (88.9) NS
OC43 7 6 (85.7) 7 (100) NS
PIV1 2 2 (100) 2 (100) NS
PIV3 9 9 (100) 9 (100) NS
Rhino 16 16 (100) 15 (93.8) NS
RSVA 7 7 (100) 6 (85.7) NS
RSVB 1 1 (100) 1 (100) NS
Total 87 86 (98.9) 75 (86.2) <0.001

Abbreviations: RT-PCR, real-time polymerase chain reaction; Adeno, adenovirus; 229E, coronavirus; BoV, bocavirus; ENT, enterovirus; INFA, inflenza virus A; MPV, metapneumovirus; OC43, coronavirus OC43; PIV1 and PIV3, parainflenza virus 1 and 3; Rhino, rhinovirus; RSVA and RSVB, respiratory syncytial virus A and B; NS, not significant.

Table 2.
Distribution of RVs in clinical nasopharyngeal specimens by AdvanSure RV real-time RT-PCR and AdvanSure RV-plus real-time RT-PCR
Target viruses No. of true positive No. (%) of positive
Overall
Single virus
Two viruses
Three viruses
AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR
Adeno 11 11 (100) 10 (90.9) 5 (5.8) 7 (8.1) 4 (4.6) 2 (2.3) 2 (2.3) 1 (1.2)
229E 2 2 (100)       1 (1.2)   1 (1.2)  
BoV 11 11 (100) 10 (90.9) 6 (5.8) 7 (8.1) 2 (2.3) 2 (2.3) 3 (3.5) 1 (1.2)
ENT 5 5 (100)       2 (2.3)   3 (3.5)  
INFA 7 7 (100) 7 (100) 7 (8.1) 7 (8.1)        
MPV 9 9 (100) 8 (88.9) 8 (9.2) 8 (9.2) 1 (1.2)      
OC43 7 6 (85.7) 7 (100) 5 (5.8) 7 (8.1) 1 (1.2)      
PIV1 2 2 (100) 2 (100) 2 (2.3) 2 (2.3)        
PIV3 9 9 (100) 9 (100) 4 (4.6) 4 (4.6) 4 (4.6) 5 (5.8) 1 (1.2)  
Rhino 16 16 (100) 15 (93.8) 5 (5.8) 7 (8.1) 7 (8.1) 7 (8.1) 4 (4.6) 1 (1.2)
RSVA 7 7 (100) 6 (85.7) 6 (6.9) 6 (6.9) 1 (1.2)      
RSVB 1 1 (100) 1 (100) 1 (1.2) 1 (1.2)        
Total 87 86 (98.9) 75 (86.2) 49 (56.3) 56 (64.4) 23 (26.4) 16 (18.4) 14 (16.1) 3 (3.5)

Abbreviations: See Table 1.

Table 3.
Detection difference in coinfections clinical nasopharyngeal swab specimens
Number AdvanSure RV-plus real-time RT-PCR AdvanSure RV real-time RT-PCR
1 Adenovirus, Rhinovirus Adenovirus
2 Bocavirus, Coronavirus Coronavirus
3 Rhinovirus, Enterovirus Rhinovirus
4 Adenovirus, Respiratory syncytial virus A, Coronavirus Adenovirus
5 Bocavirus, Rhinovirus, Enterovirus Bocavirus, Rhinovirus
6 Parainflenza Virus 3, PIV3, Coronavirus OC43 Parainflenza Virus 3, PIV3
7 Bocavirus, Adenovirus Bocavirus
8 Bocavirus, Rhinovirus, Enterovirus Bocavirus, Rhinovirus
9 Parainflenza Virus 3, PIV3, Rhinovirus, Enterovirus Parainflenza Virus 3, PIV3, Rhinovirus
10 Rhinovirus, Enterovirus Rhinovirus
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