Comparison of Culture, Direct Immunofluorescence Assay, and Multiplex Reverse Transcriptase PCR for Detection of Respiratory Viruses

Kui Hyun Yoon; Ji Hyun Cho

doi:10.3343/lmo.2011.1.4.8

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Yoon and Cho: Comparison of Culture, Direct Immunofluorescence Assay, and Multiplex Reverse Transcriptase PCR for Detection of Respiratory Viruses

Original Article

Clinical Microbiology

Laboratory Medicine Online

Laboratory Medicine Online 2011; 1(4): 221-226.

Published online: 7 October 2011

DOI: https://doi.org/10.3343/lmo.2011.1.4.8

Comparison of Culture, Direct Immunofluorescence Assay, and Multiplex Reverse Transcriptase PCR for Detection of Respiratory Viruses

Kui Hyun Yoon, M.D.¹, Ji Hyun Cho, M.D.²

¹Department of Laboratory Medicine, Wonkwang University Sanbon Hospital, Gunpo, Korea.

²Department of Laboratory Medicine, College of Medicine, Wonkwang University, Iksan, Korea.

Corresponding author: Kui Hyun Yoon. Department of Laboratory Medicine, Wonkwang University Sanbon Hospital, 1142 Sanbon-dong, Gunpo 435-040, Korea. Tel: +82-31-390-1861, Fax: +82-31-391-2085, wooju67@paran.com

Received 27 July 2011 Revised 2 September 2011 Accepted 19 September 2011

(open-access):

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

Rapid detection of causative viruses is important for the management of acute respiratory illnesses. To increase the detection rate and decrease the turnaround time (TAT) and cost, we used 24-well plates instead of R-mix shell vials and changes the report time from once on day 3 to twice on days 1 and 5 of culture. The detection rate and TATs of each culture method, direct immunofluorescence assay (DFA), and multiplex reverse transcriptase polymerase chain reaction (mPCR) were compared.

Methods

Among 2,062 nasopharyngeal swabs (NPSs) received from January 2009 to January 2011, 707 NPSs were cultured in R-mix shell vials and 1,355 NPSs were cultured in 24-well plates. We analyzed 538 NPSs simultaneously using DFA, mPCR, and culture and compared the detection rate for 7 viruses (adenovirus, influenza A and B virus; parainfluenza virus 1, 2, and 3; and respiratory syncytial virus [RSV]).

Results

The detection rate when using shell vials was 28.4% (201/707) and was 29.4% (399/1,355) on day 1 and 33.3% (452/1,355) on day 5 when using 24-well plates. In addition, of the 53 viruses that were detected on day 5, 34 were adenovirus, 7 were parainfluenza virus, 4 were influenza A virus, 3 were influenza B virus, 4 were RSV, and 1 was a mix of influenza B and parainfluenza virus. The TAT when using shell vials and 24-well plates was 4.8 days and 2.5 days, respectively. The detection rate for the 7 respiratory viruses using culture, DFA, and mPCR was 24.3%, 20.8%, and 38.5%, and the TAT was 3.7 days, 1.0 day, and 1.4 days, respectively.

Conclusions

Using 24-well plates for virus culture is an efficient method for the detection of respiratory viruses.

Keywords: respiratory viruses, virus culture, immunofluorescence assay, multiplex PCR

Figures and Tables

Table 1

Comparison of shell vial and 24-well plate culture methods for the detection of 7 major respiratory viruses in 2,062 nasopharyngeal swabs

^*P value=0.0234, ^†P value<0.0001.

Abbreviations: ADV, adenovirus; Flu-A, influenza A; Flu-B, influenza B; PIV1, parainfluenza virus 1; PIV2, parainfluenza virus 2; PIV3, parainfluenza virus 3; RSV, respiratory syncytial virus; TAT, turnaround time.

Table 2

Comparison of detection rate and turnaround time of culture, direct immunofluorescence assay, and multiplex reverse transcriptase polymerase chain reaction for the detection of 7 respiratory viruses in 538 nasopharyngeal swabs tested simultaneously

^*P value=0.0001, ^†P value<0.0001.

Abbreviations: TAT, turnaround time; DFA, direct immunofluorescence assay; mPCR, multiplex reverse transcriptase polymerase chain reaction.

Table 3

Detection of respiratory viruses by culture, direct immunofluorescence assay (DFA), and multiplex reverse transcriptase polymerase chain reaction (mPCR) in 538 nasopharyngeal swabs tested simultaneously

^*mismatch result by culture/DFA and mPCR.

Abbreviations: ADV, adenovirus; Flu-A, influenza A virus; Flu-B, influenza B virus; PIV1, parainfluenza virus 1; PIV2, parainfluenza virus 2; PIV3, parainfluenza virus 3; RSV, respiratory syncytial virus; TAT, turnaround time; Neg, negative; NG, no growth; Other, other than the 7 major respiratory viruses.

Notes

This paper was supported by Wonkwang University in 2010.

This article is available from http://www.labmedonline.org

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