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Journal of Clinical Microbiology, July 2008, p. 2298-2304, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00490-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid, Novel, Specific, High-Throughput Assay for Diagnosis of Loa loa Infection{triangledown}

Peter D. Burbelo,1 Roshan Ramanathan,2 Amy D. Klion,2 Michael J. Iadarola,1 and Thomas B. Nutman2*

Neurobiology and Pain Therapeutics Section, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research,1 Laboratory of Parasitic Diseases, National Institutes of Health, 4 Center Drive, Bethesda, Maryland 208922

Received 12 March 2008/ Returned for modification 7 May 2008/ Accepted 15 May 2008

The ability to diagnose Loa loa infection readily and accurately remains a demanding task. Among the available diagnostic methods, many are impractical for point-of-care field testing. To investigate whether luciferase immunoprecipitation systems (LIPS) can be used for rapid and specific diagnosis of L. loa infection, a LIPS assay was developed based on immunoglobulin G (IgG) and IgG4 subclass antibodies to a recombinant L. loa SXP-1 (designated LlSXP-1) antigen and tested with sera from healthy controls or patients with proven infection with L. loa, Mansonella perstans, Onchocerca volvulus, Strongyloides stercoralis, or Wuchereria bancrofti. A LIPS test measuring IgG antibody against LlSXP-1 readily differentiated L. loa-infected from uninfected patients and demonstrated markedly improved sensitivity and specificity compared with an LlSXP-1 IgG4-based enzyme-linked immunosorbent assay (67% sensitivity and 99% specificity). No significant immunoreactivity was observed with S. stercoralis-infected sera, but a small number of patients infected with O. volvulus, M. perstans, or W. bancrofti showed positive immunoreactivity. Measuring anti-IgG4-specific antibodies to LlSXP-1 showed a significant correlation (r ~ 0.85; P < 0.00001) with the anti-IgG results but showed no advantage over measuring the total IgG response alone. In contrast, a rapid LIPS format (called QLIPS) in which the tests are performed in less than 15 minutes under nonequilibrium conditions significantly improved the specificity for cross-reactive O. volvulus patient sera (100% sensitivity and 100% specificity). These results suggest that LIPS (and the even more rapid test QLIPS) represents a major advance in the ability to diagnose L. loa infection and may have future applications for point-of-care diagnostics.

* Corresponding author. Mailing address: Building 4, Room B1-03, 4 Center Drive, National Institutes of Health, Bethesda, MD 20892. Phone: (301) 496-5398. Fax: (301) 480-3757. E-mail: tnutman{at}niaid.nih.gov

{triangledown} Published ahead of print on 28 May 2008.

Journal of Clinical Microbiology, July 2008, p. 2298-2304, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00490-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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