Lab Med Online.  2011 Oct;1(4):221-226. 10.3343/lmo.2011.1.4.8.

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

Affiliations
  • 1Department of Laboratory Medicine, Wonkwang University Sanbon Hospital, Gunpo, Korea. wooju67@paran.com
  • 2Department of Laboratory Medicine, College of Medicine, Wonkwang University, Iksan, Korea.

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.

Keyword

respiratory viruses; virus culture; immunofluorescence assay; multiplex PCR
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