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. 2000 Mar;38(3):1191-5.
doi: 10.1128/JCM.38.3.1191-1195.2000.

Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR

J C Aguilar  1 , M P Pérez-Breña, M L García, N Cruz, D D Erdman, J E Echevarría
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Free PMC article

Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR

J C Aguilar et al. J Clin Microbiol. 2000 Mar.
Free PMC article

Erratum in

  • J Clin Microbiol 2000 Jul;38(7):2805

Abstract

We describe a multiplex reverse transcription-PCR (m-RT-PCR) assay that is able to detect and differentiate all known human parainfluenza viruses (HPIVs). Serial dilution experiments with reference strains that compared cell culture isolation and m-RT-PCR showed sensitivities ranging from 0.0004 50% tissue culture infective dose (TCID(50)) for HPIV type 4B (HPIV-4B) to 32 TCID(50)s for HPIV-3. As few as 10 plasmids containing HPIV PCR products could be detected in all cases. When 201 nasopharyngeal aspirate specimens from pediatric patients hospitalized for lower respiratory illness were tested, m-RT-PCR assay detected 64 HPIVs (24 HPIV-3, 23 HPIV-1, 10 HPIV-4, and 7 HPIV-2), while only 42 of them (21 HPIV-1, 14 HPIV-3, 6 HPIV-2, and 1 HPIV-4 isolates) grew in cell culture. Our m-RT-PCR assay was more sensitive than either cell culture isolation or indirect immunofluorescence with monoclonal antibodies for the detection of HPIV infections. Also, HPIV-4 was more frequently detected than HPIV-2 in this study, suggesting that it may have been underestimated as a lower respiratory tract pathogen because of the insensitivity of cell culture.

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Figures

FIG. 1
FIG. 1
Typing of HPIVs and subtyping of HPIV-4 by m-RT-PCR: mixture of HPIVs (lane 1), HPIV-1 (lane 2), HPIV-4 (lane 3), HPIV-2 (lane 4), HPIV-3 (lane 5), negative control (lane 6), marker (DNA molecular weight marker VIII; Boehringer Mannheim) (lane 7), HPIV-4A (lane 8), and HPIV-4B (lane 9).
FIG. 2
FIG. 2
Comparison of the temporal distribution between culture-positive (formula image) and m-RT-PCR-positive (■) results. Dic., December; En., January.
See this image and copyright information in PMC

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