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Journal of Clinical Microbiology, May 2001, p. 1696-1701, Vol. 39, No. 5
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.5.1696-1701.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Evaluation of the Hexaplex Assay for Detection
of Respiratory Viruses in Children
Sue C.
Kehl,1,2,*
Kelly J.
Henrickson,2,3,4
Weimin
Hua,4 and
Jiang
Fan3
Departments of
Pathology1 and
Pediatrics,3 Medical College of
Wisconsin, Children's Hospital of
Wisconsin,2 and Prodesse,
Inc.,4 Milwaukee, Wisconsin
Received 2 June 2000/Returned for modification 21 August
2000/Accepted 23 February 2001
The Hexaplex assay (Prodesse, Inc., Milwaukee, Wis.) is a multiplex
reverse transcriptase (RT)-PCR assay for the detection of parainfluenza
virus types 1, 2, and 3, respiratory syncytial virus (RSV) types A and
B, and influenza virus types A and B. We evaluated the Hexaplex assay
in comparison with conventional viral cell cultures and rapid enzyme
immunoassays (EIAs) for RSV (Directigen; Becton Dickinson Inc.,
Cockeysville, Md.) and influenza A virus (Abbott Test Pack; Abbott
Laboratories, Abbott Park, Ill.) for the detection of respiratory
viruses from pediatric respiratory specimens obtained from children
seen at Children's Hospital of Wisconsin from December 1997 through
May 1998. A total of 363 respiratory specimens were evaluated. The
tissue culture prevalence of parainfluenza virus during this period of
time was low (1.1%). The sensitivity, specificity, and positive and
negative predictive value of Hexaplex compared to tissue culture for
the detection of parainfluenza virus were 100, 95.8, 19.0, and 100%,
respectively. Only one specimen was determined to contain influenza B
virus by Hexaplex; it was tissue culture negative. A specimen was
considered to contain RSV or influenza A virus when it was either
culture positive or culture negative but Hexaplex and EIA positive.
Prior to the analysis of discrepant results, the sensitivity,
specificity, and positive and negative predictive value for the
detection of RSV were 91.2, 100, 100, and 98.0%, respectively, for
tissue culture; 84.5, 100, 100, and 96.6% for EIA; and 98.5, 91.5, 72.8, and 99.6% for Hexaplex, respectively. The sensitivity,
specificity, and positive and negative predictive value for the
detection of influenza A virus prior to the analysis of discrepant
results were 100, 100, 100, and 100%, respectively, for culture, 78.0, 100, 100, and 89.4% for EIA, respectively, and 95.1, 94.1, 67.2, and
99.3% for Hexaplex, respectively. Culture- and/or EIA-negative,
Hexaplex-positive specimens were analyzed by a second RT-PCR assay
which used primers specific for a different genomic region than that
used in the Hexaplex assay. After analysis of these discrepant results,
the sensitivity, specificity, and positive and negative predictive value for the detection of RSV were 74.3, 100, 100, and 93.5%, respectively, for tissue culture; 70.3, 100, 100, and 92.5% for EIA;
and 98.6, 97.4, 91.2, and 99.6% for Hexaplex. The sensitivity, specificity, and positive and negative predictive value for the detection of influenza A virus were 83.3, 100, 100, and 97.4%, respectively, for tissue culture; 69.4, 100, 100, and 83.3% for EIA;
and 95.8, 98.7, 92.0, and 99.3% for Hexaplex. Hexaplex is a rapid,
sensitive, and specific method for the detection of the seven most
common respiratory viruses in children.
*
Corresponding author. Mailing address: Department of
Pathology, Medical College of Wisconsin, P.O. Box 26509, Milwaukee, WI 53226-0509. Phone: (414) 266-2529. Fax: (414) 266-2779. E-mail: kskehl{at}mcw.edu.
Journal of Clinical Microbiology, May 2001, p. 1696-1701, Vol. 39, No. 5
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.5.1696-1701.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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Copyright © 2001 by the American Society for Microbiology. All rights reserved.