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Journal of Clinical Microbiology, December 2004, p. 5596-5603, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5596-5603.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Practical Implementation of a Multiplex PCR for Acute Respiratory Tract Infections in Children

Paul Gruteke,1* Afina S. Glas,2 Mirjam Dierdorp,1 Willem B. Vreede,3 Jan-Willem Pilon,4 and Sylvia M. Bruisten1

Municipal Public Health Laboratory, Municipal Health Service,1 Clinical Epidemiology and Biostatistics, Academic Medical Center, Amsterdam,2 Flevo Hospital, Almere,3 IJsselmeer Hospitals, Lelystad and Emmeloord, The Netherlands4

Received 23 December 2003/ Returned for modification 20 April 2004/ Accepted 27 July 2004

Molecular testing for acute respiratory infections (ARIs) has documented value but limited implementation due to questions that typically slow the acceptance of new tests. This study sought to address these questions and achieve implementation. Rhinovirus was added to a nested multiplex PCR (M-PCR), increasing its diagnostic yield. Over one winter, three hospital pediatric departments used the M-PCR to complement their direct fluorescent-antibody assay (DFA) for respiratory syncytial virus (RSV). Clinicians recorded "pretest probability estimates" (using continuous scales for various pathogen groups) for comparison with test results; treatments and test turnaround times were also recorded. Transnasal and throat swabs, with or without nasopharyngeal aspirate (NPA), were M-PCR tested. NPA-containing sample sets found to be RSV positive by DFA were not further tested. Single PCR for human metapneumovirus (hMPV) was performed retrospectively. Of 178 ARI episodes representing 172 patients, NPA was included in 97 sample sets; 54 (56%) were determined to be RSV positive. The other NPA-containing sample sets (n = 43) yielded 27 findings (63%), and the swab-only sets (n = 81) yielded 47 findings (58%); rhinovirus was found most often. Testing for hMPV yielded seven positive results. M-PCR median turnaround times were 4 days in swab-only samples and 5 days with NPA. Antibiotics were prescribed in 50 episodes, at rates similar for RSV and rhinovirus. Pretest probability estimates of a viral cause were lower in episodes caused by rhinovirus than in episodes caused by RSV. The hospitals continued to use M-PCR for NPA-containing samples found to be RSV negative by DFA. Test implementation is more likely with higher diagnostic yield and a protocol that reflects day-to-day clinical and laboratory operations.

* Corresponding author. Mailing address: Municipal Public Health Laboratory Amsterdam, Nieuwe Achtergracht 100, P.O. Box 2200, 1000 CE Amsterdam, The Netherlands. Phone: 31-20-5555-275. Fax: 31-20-5555-629. E-mail: pgruteke{at}gggd.amsterdam.nl.

Journal of Clinical Microbiology, December 2004, p. 5596-5603, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5596-5603.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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