Korean J Lab Med.  2010 Jun;30(3):289-294. 10.3343/kjlm.2010.30.3.289.

Comparison of R-mix Virus Culture and Multiplex Reverse Transcriptase-PCR for the Rapid Detection of Respiratory Viruses

• Lim G
• Park TS
• Suh JT
• Lee HJ

• KMID: 1781629
• DOI: http://doi.org/10.3343/kjlm.2010.30.3.289

Abstract

BACKGROUND
Respiratory viral infections can become epidemic due to high contagiosity. Since there was no rapid diagnostic method for complete diagnosis in the past, diagnosis was solely made on the basis of clinical symptoms or the time of infection. With recent developments in rapid diagnostic methods like multiplex reverse transcriptase (RT)-PCR, R-mix virus culture, etc., early detection and effective treatment of respiratory viral infections is possible. Herein, we compared the efficiency of multiplex RT-PCR and the R-mix virus culture for the rapid detection of respiratory viruses.
METHODS
We used 96 nasopharyngeal swab specimens for culturing respiratory viruses using R-mix (Diagnostics Hybrids Inc., USA). Afterwards, multiplex RT-PCR was performed using specimens stored at -70degrees C.
RESULTS
R-mix virus culture yielded positive results in 34 cases (35.4%) and multiplex RT-PCR in 73 cases (76.0%). Both methods yielded identical results in 51 cases (29 positive cases and 22 negative cases). Among 45 cases that showed different results, 40 showed negative results in R-mix virus culture and positive results in multiplex RT-PCR, and 1 showed positive result in R-mix virus culture and negative result in multiplex RT-PCR. Different viruses were detected in the remaining 4 cases by both the methods.
CONCLUSIONS
Multiplex RT-PCR provided faster results and had higher detection rates than R-mix virus culture. Further, unlike R-mix virus culture, multiplex RT-PCR can be used to identify new respiratory viruses. Therefore, multiplex RT-PCR is more useful than R-mix virus culture in the diagnosis of respiratory virus infection.

Keyword

R-mix virus culture; Respiratory virus detection; Multiplex respiratory RT-PCR

MeSH Terms

Child
Child, Preschool
Female
Humans
Infant
Infant, Newborn
Male
Middle Aged
RNA, Viral/analysis
Reagent Kits, Diagnostic
Respiratory Tract Infections/*virology
Retrospective Studies
*Reverse Transcriptase Polymerase Chain Reaction
*Virus Cultivation
Virus Diseases/*diagnosis
Viruses/genetics/*isolation &purification

Reference

1.Kim MR., Lee HR., Lee GM. Epidemiology of acute viral respiratory tract infections in Korean children. J Infect. 2000. 41:152–8.

2.Kim SH., Huh JH., Bae SY., Kim JS., Yoon SY., Lim CS, et al. Epidemiology of respiratory viral infection in 2004-2006. Korean J Lab Med. 2006. 26:351–7. (김선형, 허지훈, 배숙영, 김장수, 윤수영, 임채승등. 2004-2006년의 호흡기 바이러스 감염의 역학. 대한진단검사의학회지 2006;26:351-7.).

3.Ginocchio CC. Detection of respiratory viruses using non-molecular based methods. J Clin Virol. 2007. 40(S1):11–4.

4.Leland DS., Ginocchio CC. Role of cell culture for virus detection in the age of technology. Clin Microbiol Rev. 2007. 20:49–78.

5.McPherson RA, Pincus MR, editors. Henry's Clinical diagnosis and management by laboratory methods. 21st ed.Philadelphia: Saunders;2007. p. 975–84.

6.Barenfanger J., Drake C., Mueller T., Troutt T., O'Brien J., Guttman K. R-Mix cells are faster, at least as sensitive and marginally more costly than conventional cell lines for the detection of respiratory viruses. J Clin Virol. 2001. 22:101–10.

7.LaSala PR., Bufton KK., Ismail N., Smith MB. Prospective comparison of R-mix shell vial system with direct antigen tests and conventional cell culture for respiratory virus detection. J Clin Virol. 2007. 38:210–6.

8.St George K., Patel NM., Hartwig RA., Scholl DR., Jollick JA Jr., Kauffmann LM, et al. Rapid and sensitive detection of respiratory virus infections for directed antiviral treatment using R-Mix cultures. J Clin Virol. 2002. 24:107–15.

9.Mahony JB. Detection of respiratory viruses by molecular methods. Clin Microbiol Rev. 2008. 21:716–47.

10.Yoo SJ., Kuak EY., Shin BM. Detection of 12 respiratory viruses with two-set multiplex reverse transcriptase-PCR assay using a dual priming oligonucleotide system. Korean J Lab Med. 2007. 27:420–7. (유수진, 곽은영, 신보문. Dual priming oligonucleotide system을이용한다중 역전사 PCR 키트를 통한 12가지 호흡기 바이러스의 검출. 대한진단검사의학회지 2007;27:420-7.).

11.Lin TY., Huang YC., Ning HC., Tsao KC. Surveillance of respiratory viral infections among pediatric outpatients in northern Taiwan. J Clin Virol. 2004. 30:81–5.

12.Maitreyi RS., Broor S., Kabra SK., Ghosh M., Seth P., Dar L, et al. Rapid detection of respiratory viruses by centrifugation enhanced cultures from children with acute lower respiratory tract infections. J Clin Virol. 2000. 16:41–7.

13.Fauci AS., Kasper DL. .Harrison's principles of internal medicine. 17th ed.Boston: McGraw-Hill;2008. p. 1120–32.

14.Ahn HS. Pediatrics. 9th ed.Seoul: Daehan;2008. :629-53. (안효섭편. 소아과학. 9판. 서울: 대한교과서,. 2008:p. 629–53.

15.Kim SR., Ki CS., Lee NY. Rapid detection and identification of 12 respiratory viruses using a dual priming oligonucleotide system-based multiplex PCR assay. J Virol Methods. 2009. 156:111–6.

16.Kang JO., Kim EC., Lee KM., Lee NY., Lee CG. Surveillance for respiratory virus testing situation in Korea and epidemiology for the respiratory viruses detected in 5 university hospitals. Korean J Clin Microbiol. 2007. 10:102–8. (강정옥, 김의종, 이규만, 이남용, 이창규. 국내의료기관의 호흡기 바이러스 검사현황 및 5개 대학병원에서 검출된 호흡기바이러스역학. 대한임상미생물학회지 2007;10:102-8.).

17.Yoon KH., Cho JH. Detection of respiratory viruses in children by multiplex reverse transcriptase PCR, direct immunofluorescence assay, and shell vial culture. Korean J Clin Microbiol. 2009. 12:110–5. (윤귀현 및 조지현. 소아에서 다중 역전사효소 PCR법, 직접면역형광법,쉘바이알배양법을 이용한 호흡기 바이러스의 검출. 대한임상미생물학회지 2009;12:110-5.).

18.Roh KH., Kim J., Nam MH., Yoon S., Lee CK., Lee K, et al. Comparison of the Seeplex reverse transcription PCR assay with the R-mix viral culture and immunofluorescence techniques for detection of eight respiratory viruses. Ann Clin Lab Sci. 2008. 38:41–6.

19.Kim KH., Lee JH., Sun DS., Kim YB., Choi YJ., Park JS, et al. Detection and clinical manifestations of twelve respiratory viruses in hospitalized children with acute lower respiratory tract infections: focus on human metapneumovirus, human rhinovirus and human coronavirus. Korean J Pediatr. 2008. 51:834–41. (김금향, 이정호, 선동신, 김용배, 최영진, 박준수등. 하기도감염으로입원한소아에서12종 바이러스의 검출 및 임상 양상 Korean J Pediatr 2008;51:834-41.).

20.Canducci F., Debiaggi M., Sampaolo M., Marinozzi MC., Berre S., Terulla C, et al. Two-year prospective study of single infections and coinfections by respiratory syncytial virus and viruses identified recently in infants with acute respiratory disease. J Med Virol. 2008. 80:716–23.

21.Sung H., Park SJ., Woo YD., Choi BH., Kim MN. Evaluation of Seeplex RV detection kit for detecting rhinovirus, human metapneumovirus, and coronavirus. Korean J Lab Med. 2008. 28:109–17. (성흥섭, 박숙자,우영대, 최병후, 김미나. Rhinovirus, Human Metapneumovirus, Coronavirus 검출을위한 Seeplex™ RV Detection 키트의평가. 대한진단검사의학회지 2008;28:109-17.).

22.Fouchier RA., Rimmelzwaan GF., Kuiken T., Osterhaus AD. Newer respiratory virus infections: human metapneumovirus, avian influenza virus, and human coronaviruses. Curr Opin Infect Dis. 2005. 18:141–6.