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Detection of precytopathic effect of enteroviruses in clinical specimens by centrifugation-enhanced antigen detection
S M Lipson1, K David, F Shaikh
1Department of Biomedical Sciences, Long Island University, Brookville, New York, USA. montmor@aol.com
Abstract:
Rapid enterovirus detection is important for decisions about antibiotic administration and length of hospital stay. The efficacy of rapid antigen detection-cell culture amplification (Ag-CCA) was evaluated with monoclonal antibodies (MAbs) 5-D8/1 (DAKO) and Pan-Enterovirus clone 2E11 (Chemicon) with 10 poliovirus, echovirus, and coxsackievirus type A and B stock isolates and College of American Pathologists check samples. By using Ag-CCA technology, MAb 2E11 was more sensitive than 5-D8/1 at detecting a greater number of stock isolates at or past tube (cytopathic effect [CPE]) culture (TC) end points. The efficacy of Ag-CCA in the clinical setting was subsequently confirmed with 273 consecutively freshly collected nasopharyngeal aspirate or swab specimens, rectal swab, and cerebrospinal fluid specimens during the 1999 enterovirus season. All specimens were tested by Ag-CCA in parallel with rhesus monkey kidney (RhMk), MRC-5, and A549 conventional TCs. Approximately 60% of field specimens were additionally tested with Hep-2 and HNK conventional TCs. Sixty-two percent of the clinical specimens tested were Ag-CCA positive after 48 h. Among 51 isolates, the mean time to CPE or culture confirmation was 5.5 days (range, 2 to 18 days). After 48 h, Ag-CCA achieved sensitivity, specificity, and positive and negative predictive values of 62, 100, 100, and 93%, respectively. During the same period, TC-CPE displayed test parameters of 12, 100, 100, and 85%, respectively. After 5 days, the sensitivity and specificity of Ag-CCA increased to 92 and 98%, respectively. Within the same period, isolation attained sensitivity and specificity of 52 and 100%, respectively. Although Ag-CCA displayed slightly reduced sensitivity and reduced specificity compared with conventional cell culture after 14 days, the markedly superior 48-h enterovirus Ag-CCA detection rate supports incorporation of this assay into the routine clinical setting.
Insights
Rapid antigen detection-cell culture amplification (Ag-CCA) offers faster enterovirus detection than traditional methods. This rapid test aids clinical decisions by providing quicker results for patient management.
Area of Science:
- Virology
- Clinical Diagnostics
- Infectious Diseases
Background:
- Timely enterovirus detection is crucial for guiding antibiotic use and hospital stay duration.
- Conventional cell culture methods for enterovirus diagnosis can be time-consuming.
- Evaluating rapid diagnostic assays is essential for improving clinical workflow.
Purpose of the Study:
- To assess the efficacy of rapid antigen detection-cell culture amplification (Ag-CCA) for enterovirus detection.
- To compare the performance of two monoclonal antibodies (MAbs) 5-D8/1 and Pan-Enterovirus clone 2E11 within the Ag-CCA assay.
- To validate the clinical utility of Ag-CCA in nasopharyngeal, rectal, and cerebrospinal fluid specimens.
Main Methods:
- Ag-CCA assay utilizing MAbs 5-D8/1 and Pan-Enterovirus clone 2E11 was tested with enterovirus stock isolates.
- Clinical specimens (nasopharyngeal aspirates/swabs, rectal swabs, CSF) were tested using Ag-CCA in parallel with conventional cell cultures (RhMK, MRC-5, A549, Hep-2, HNK).
- Performance metrics including sensitivity, specificity, and predictive values were calculated at different time points.
Main Results:
- MAb 2E11 demonstrated higher sensitivity than 5-D8/1 in detecting stock isolates.
- Ag-CCA detected enteroviruses in 62% of clinical specimens within 48 hours, with high specificity (100%) and negative predictive value (93%).
- After 5 days, Ag-CCA sensitivity reached 92%, surpassing conventional isolation rates (52%).
Conclusions:
- The Ag-CCA assay, particularly with MAb 2E11, provides rapid and reliable enterovirus detection.
- Ag-CCA significantly reduces the time to diagnosis compared to conventional cell culture methods.
- The study supports the integration of Ag-CCA into routine clinical settings for efficient enterovirus management.
