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Clinical and Diagnostic Laboratory Immunology, February 2005, p. 235-241, Vol. 12, No. 2
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.2.235-241.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Quantitative PCR-Enhanced Immunoassay for Measurement of Enteroviral Immunoglobulin M Antibody and Diagnosis of Aseptic Meningitis

Amal Elfaitouri,1 Nahla Mohamed,1 Jan Fohlman,2 Robert Aspholm,1 Gun Frisk,3 Göran Friman,2 Lars Magnius,1,4 and Jonas Blomberg1*

Section of Virology,1 Section of Infectious Diseases, Department of Medical Sciences,2 Section of Pediatrics, Uppsala University Hospital, Uppsala,3 Virology Unit, Swedish Institute for Infectious Disease Control, Solna, Sweden4

Received 5 January 2004/ Returned for modification 4 May 2004/ Accepted 4 November 2004

A PCR-enhanced immunoassay (PIA) to detect enterovirus (EV) immunoglobulin M (IgM) for diagnosis of recent EV infection was recently developed. This test was compared with another EV IgM capture technique, the solid-phase reverse immunosorbent test (SPRIST). Fourteen of 43 serum samples from aseptic meningitis patients were positive by PIA, whereas 10 were positive by SPRIST. One of 39 control serum samples was weakly positive by PIA. A single-serum-dilution real-time PCR-based PIA for EV IgM (quantitative PIA [QPIA]) was also developed and evaluated against PIA, SPRIST, an EV IgM radioimmunoassay (RIA), and clinical data. A mixture of 12 EVs was used as the antigen. Results from investigating four groups of serum samples were as follows. (i) The nine PIA-positive serum samples in group 1 were all positive by QPIA. (ii) Group 2 consisted of 59 serum samples from aseptic meningitis patients. Nineteen of 30 serum samples (63%) taken at hospital admission were positive by QPIA. Of these, 17 were positive in EV PCR. (iii) None of the 30 control serum samples in group 3 were positive by QPIA. (iv) For the 24 serum samples in group 4, of which 11 were positive and 13 were negative by RIA, the QPIA results were completely concordant. The sensitivity and specificity of QPIA for diagnosis of EV infection were 70 and 80%, respectively. QPIA provides a rational strategy for the detection of EV IgM, allows the use of viral antigens with minimal purification, and needs no virus-specific reagents apart from those in the PCR. QPIA is a generally applicable method for the detection of viral IgM in IgM capture assays.

* Corresponding author. Mailing address: Section of Clinical Virology, Department of Medical Sciences, Academic Hospital, S-751 85 Uppsala, Sweden. Phone: 46 18 611 55 93. Fax: 46 18 55 10 12. E-mail: jonas.blomberg{at}kvir.uu.se.

Clinical and Diagnostic Laboratory Immunology, February 2005, p. 235-241, Vol. 12, No. 2
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.2.235-241.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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