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

Gayoung Lim; Tae Sung Park; Jin-Tae Suh; Hee Joo Lee

doi:10.3343/kjlm.2010.30.3.289

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Lim, Park, Suh, and Lee: Comparison of R-mix Virus Culture and Multiplex Reverse Transcriptase-PCR for the Rapid Detection of Respiratory Viruses

Original Article

Korean J Leg Med 2010;30(3):289-294.

Published online: 14 January 2010

DOI: https://doi.org/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

Gayoung Lim, M.D., Tae Sung Park, M.D., Jin-Tae Suh, M.D., Hee Joo Lee, M.D.

Department of Laboratory Medicine, Kyung Hee University, College of Medicine, Seoul, Korea

Corresponding author: Hee Joo Lee, M.D. Department of Laboratory Medicine, Kyung Hee University School of Medicine, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-702, Korea Tel: +82-2-958-8672, Fax: +82-2-958-8609 E-mail: leehejo@khmc.or.kr

Revised 14 October 20097 April 2010 Accepted 3 May 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 −70°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.

Keywords: R-mix virus culture, Respiratory virus detection, Multiplex respiratory RT-PCR

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Table 1.

Clinical characteristics of patients

Characteristics	N of patients (%)
Total N of patients	96
Sex (male/female)	56/40
Age (yr)
0-1	66 (68.8)
2-5	22 (22.9)
6-11	7 (7.3)
59	1 (1.0)
Diagnosis
Upper respiratory infection	15 (15.6)
Acute pharyngitis	8 (8.3)
Acute tonsilitis	3 (3.1)
Acute otitis media	1 (1.0)
Not available	3 (3.1)
Lower respiratory infection	74 (77.1)
Pneumonia	48 (50.0)
Acute bronchiolitis	17 (17.7)
Croup	9 (9.4)
Others∗	7 (7.3)

^∗ Acute gastric enteritis, 4; Urinary tract infection, 2; rule out sepsis, 1.

Table 2.

Comparison between the results of R-mix culture and multiplex RT-PCR

Multiplex RT-PCR	R-mix culture		Total (%)
Multiplex RT-PCR	Positive (%)	Negative (%)	Total (%)
Positive∗	33^† (34.3)	30^‡ (31.3)	73 (76.0)
		10^§ (10.4)
Negative	1 (1.0)	22 (22.9)	23 (24.0)
Total	34 (35.4)	62 (64.6)	96 (100)

^∗ Multiplex RT-PCR results suggested that 14 patients were co-infected.

^† 4 cases showed different results with R-mix culture and multiplex RT-PCR.

^‡ Multiplex RT-PCR analysis was positive for 8 respiratory viruses (influenza A/B, parainfluenza 1/2/3, respiratory syncytial virus type A/B adenovirus) and 4 other respiratory viruses (metapneumovirus, rhinovirus, coronavirus 229E/NL63, and corona virus OC43). ^§Multiplex RT-PCR results were positive for only 4 extra respiratory viruses (metapneumovirus, rhinovirus, coronavirus 229E/NL63, and corona virus OC43).

Table 3.

R-mix virus culture and multiplex RT-PCR results of respiratory viruses

Respiratory viruses	N of positive results (%)
Respiratory viruses	R-mix culture	Multiplex RT-PCR
Influenza A∗	16 (45.7)	25 (27.2)
Respiratory syncytial virus	14 (40.0)	31 (33.7)
Parainfluenza 1	1 (2.9)	2 (2.2)
Parainfluenza 2	2 (5.7)	0 (0.0)
Parainfluenza 3	1 (2.9)	1 (1.1)
Adenovirus	1 (2.9)	7 (7.6)
Coronavirus 229E/NL63	-	13 (14.1)
Coronavirus OC43/HKU1	-	7 (7.6)
Rhinovirus	-	5 (5.4)
Metapneumovirus	-	1 (1.1)
Total	35^† (100)	92^† (100)

^∗ Co-infection with adenovirus: 1 case.

^† Cases with co-infection are separately included in the total number of cases. R-mix virus culture, 1 positive case; multiplex RT-PCR, 14 positive cases.

Table 4.

Co-infection with respiratory viruses detected by multiplex RT-PCR

Respiratory viruses	N of patients
Corona virus 229E/NL63+RSV A	6
Corona virus 229E/NL63+influenza A	2
Corona virus 229E/NL63+adenovirus	1
Corona virus 229E/NL63+corona virus OC43/HKU1+adenovirus	1
Corona virus 229E/NL63+adenovirus+rhinovirus	1
Corona virus OC43/HKU1+RSV A+rhinovirus	1
Corona virus OC43/HKU1+adenovirus, rhinovirus+RSV A	1
Adenovirus+influenza A	1

Abbreviations: RSV, respiratory syncytial virus.

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