Detection of 11 Common Viral and Bacterial Pathogens Causing Community-Acquired Pneumonia or Sepsis in Asymptomatic Patients by Using a Multiplex Reverse Transcription-PCR Assay with Manual (Enzyme Hybridization) or Automated (Electronic Microarray) Detection
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Published:2008-09
Issue:9
Volume:46
Page:3063-3072
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ISSN:0095-1137
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Container-title:Journal of Clinical Microbiology
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language:en
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Short-container-title:J Clin Microbiol
Author:
Kumar Swati1, Wang Lihua2, Fan Jiang2, Kraft Andrea2, Bose Michael E.2, Tiwari Sagarika2, Van Dyke Meredith2, Haigis Robert3, Luo Tingquo3, Ghosh Madhushree3, Tang Huong3, Haghnia Marjan3, Mather Elizabeth L.3, Weisburg William G.3, Henrickson Kelly J.1
Affiliation:
1. Pediatric Infectious Diseases, Children's Hospital of Wisconsin, Suite C450, P.O. Box 1997, Milwaukee, Wisconsin 53201-1997 2. Department of Pediatrics, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, Wisconsin 53226 3. Nanogen, Inc., 10398 Pacific Center Court, San Diego, California 92121
Abstract
ABSTRACT
Community-acquired pneumonia (CAP) and sepsis are important causes of morbidity and mortality. We describe the development of two molecular assays for the detection of 11 common viral and bacterial agents of CAP and sepsis: influenza virus A, influenza virus B, respiratory syncytial virus A (RSV A), RSV B,
Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, Legionella micdadei, Bordetella pertussis, Staphylococcus aureus
, and
Streptococcus pneumoniae
. Further, we report the prevalence of carriage of these pathogens in respiratory, skin, and serum specimens from 243 asymptomatic children and adults. The detection of pathogens was done using both a manual enzyme hybridization assay and an automated electronic microarray following reverse transcription and PCR amplification. The analytical sensitivities ranged between 0.01 and 100 50% tissue culture infective doses, cells, or CFU per ml for both detection methods. Analytical specificity testing demonstrated no significant cross-reactivity among 19 other common respiratory organisms. One hundred spiked “surrogate” clinical specimens were all correctly identified with 100% specificity (95% confidence interval, 100%). Overall, 28 (21.7%) of 129 nasopharyngeal specimens, 11 of 100 skin specimens, and 2 of 100 serum specimens from asymptomatic subjects tested positive for one or more pathogens, with
S. pneumoniae
and
S. aureus
giving 89% of the positive results. Our data suggest that asymptomatic carriage makes the use of molecular assays problematic for the detection of
S. pneumoniae
or
S. aureus
in upper respiratory tract secretions; however, the specimens tested showed virtually no carriage of the other nine viral and bacterial pathogens, and the detection of these pathogens should not be a significant diagnostic problem. In addition, slightly less sensitive molecular assays may have better correlation with clinical disease in the case of CAP.
Publisher
American Society for Microbiology
Subject
Microbiology (medical)
Reference43 articles.
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