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. 2016 Jun;32(6):612-9.
doi: 10.1089/aid.2014.0351. Epub 2016 Mar 15.

Novel Time-Resolved Fluorescence Europium Nanoparticle Immunoassay for Detection of Human Immunodeficiency Virus-1 Group O Viruses Using Microplate and Microchip Platforms

Mohan Kumar Haleyur Giri Setty  1 , Jikun Liu  1 , Prerna Mahtani  1 , Panhe Zhang  1 , Bingchen Du  1 , Viswanath Ragupathy  1 , Krishnakumar Devadas  1 , Indira K Hewlett  1
Affiliations

Novel Time-Resolved Fluorescence Europium Nanoparticle Immunoassay for Detection of Human Immunodeficiency Virus-1 Group O Viruses Using Microplate and Microchip Platforms

Mohan Kumar Haleyur Giri Setty et al. AIDS Res Hum Retroviruses. 2016 Jun.

Abstract

Accurate detection and quantification of HIV-1 group O viruses have been challenging for currently available HIV assays. We have developed a novel time-resolved fluorescence (TRF) europium nanoparticle immunoassay for HIV-1 group O detection using a conventional microplate enzyme-linked immunosorbent assay (ELISA) and a microchip platform. We screened several antibodies for optimal reactivity with several HIV-1 group O strains and identified antibodies that can detect all the strains of HIV-1 group O that were available for testing. The antibodies were used to develop a conventional ELISA format assay and an in-house developed europium nanoparticle-based assay for sensitivity. The method was evaluated on both microwell plate and microchip platforms. We identified two specific and sensitive antibodies among the six we screened. The antibodies, C65691 and ANT-152, were able to quantify 15 and detect all 17 group O viruses, respectively, as they were broadly cross-reactive with all HIV-1 group O strains and yielded better signals compared with other antibodies. We have developed a sensitive assay that reflects the actual viral load in group O samples by using an appropriate combination of p24 antibodies that enhance group O detection and a highly sensitive TRF-based europium nanoparticle for detection. The combination of ANT-152 and C65690M in the ratio 3:1 was able to give significantly higher signals in our europium-based assay compared with using any single antibody.

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Figures

<b>FIG. 1.</b>
FIG. 1.
(A) Screening of different commercially available monoclonal HIV p24 antibodies with HIV 1 group O strains on ENIA platform. (B) Comparision of monoclonal HIV p24antibodies, ANT-152 with C65690M, on ENIA platform. ENIA, europium nanoparticle immunoassay. Color images available online at www.liebertpub.com/aid
<b>FIG. 2.</b>
FIG. 2.
Comparision of HIV 1 group O quantification by Perkin Elmer Kit and ENIA for ANT-152 with C65690M antibodies. Color images available online at www.liebertpub.com/aid
<b>FIG. 3.</b>
FIG. 3.
Comparision of HIV 1 group O signals on ENIA with 3:1 and 1:3 ratios of ANT-152 with C65690M antibodies. Color images available online at www.liebertpub.com/aid
<b>FIG. 4.</b>
FIG. 4.
Detection of 5 pg of p24 in group O strains with different antibodies. Color images available online at www.liebertpub.com/aid
<b>FIG. 5.</b>
FIG. 5.
Comparison of detection of 25 pg of group O strains in phosphate buffered saline Tween 20 obtained from Thermo Scientific Product #37539 versus plasma. Color images available online at www.liebertpub.com/aid
<b>FIG. 6.</b>
FIG. 6.
HIV 1 p24 sensitivity and dynamic range curve for microchip ENIA.
<b>FIG. 7.</b>
FIG. 7.
HIV 1 group O p24 detection on microchip platform using ENIA.
See this image and copyright information in PMC

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