Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling

Akira Nakatsuma, Mugiho Kaneda, Hiromi Kodama, Mika Morikawa, Satoshi Watabe, Kazunari Nakaishi, Masakane Yamashita, Teruki Yoshimura, Toshiaki Miura, Masaki Ninomiya, Etsuro Ito

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

To reduce the window period between HIV-1 infection and the ability to diagnose it, a fourthgeneration immunoassay including the detection of HIV-1 p24 antigen has been developed. However, because the commercially available systems for this assay use special, high-cost instruments to measure, for example, chemiluminescence, it is performed only by diagnostics companies and hub hospitals. To overcome this limitation, we applied an ultrasensitive ELISA coupled with a thio-NAD cycling, which is based on a usual enzyme immunoassay without special instruments, to detect HIV-1 p24. The p24 detection limit by our ultrasensitive ELISA was 0.0065 IU/assay (i.e., ca. 10-18 moles/assay). Because HIV-1 p24 antigen is thought to be present in the virion in much greater numbers than viral RNA copies, the value of 10-18 moles of the p24/assay corresponds to ca. 103 copies of the HIV-1 RNA/assay. That is, our ultrasensitive ELISA is chasing the detection limit (102 copies/assay) obtained by PCR-based nucleic acid testing (NAT) with a margin of only one different order. Further, the detection limit by our ultrasensitive ELISA is less than that mandated for a CE-marked HIV antigen/antibody assay. An additional recovery test using blood supported the reliability of our ultrasensitive ELISA.

Original languageEnglish
Article numbere0131319
JournalPLoS One
Volume10
Issue number6
DOIs
Publication statusPublished - 2015 Jun 22
Externally publishedYes

Fingerprint

Human immunodeficiency virus 1
HIV-1
Assays
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
HIV Core Protein p24
assays
Limit of Detection
detection limit
HIV Antigens
antigens
HIV Antibodies
Viral RNA
Hematologic Tests
Luminescence
RNA
Immunoenzyme Techniques
Immunoassay
Virion
Antigens

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Nakatsuma, A., Kaneda, M., Kodama, H., Morikawa, M., Watabe, S., Nakaishi, K., ... Ito, E. (2015). Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling. PLoS One, 10(6), [e0131319]. https://doi.org/10.1371/journal.pone.0131319

Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling. / Nakatsuma, Akira; Kaneda, Mugiho; Kodama, Hiromi; Morikawa, Mika; Watabe, Satoshi; Nakaishi, Kazunari; Yamashita, Masakane; Yoshimura, Teruki; Miura, Toshiaki; Ninomiya, Masaki; Ito, Etsuro.

In: PLoS One, Vol. 10, No. 6, e0131319, 22.06.2015.

Research output: Contribution to journalArticle

Nakatsuma, A, Kaneda, M, Kodama, H, Morikawa, M, Watabe, S, Nakaishi, K, Yamashita, M, Yoshimura, T, Miura, T, Ninomiya, M & Ito, E 2015, 'Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling', PLoS One, vol. 10, no. 6, e0131319. https://doi.org/10.1371/journal.pone.0131319
Nakatsuma A, Kaneda M, Kodama H, Morikawa M, Watabe S, Nakaishi K et al. Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling. PLoS One. 2015 Jun 22;10(6). e0131319. https://doi.org/10.1371/journal.pone.0131319
Nakatsuma, Akira ; Kaneda, Mugiho ; Kodama, Hiromi ; Morikawa, Mika ; Watabe, Satoshi ; Nakaishi, Kazunari ; Yamashita, Masakane ; Yoshimura, Teruki ; Miura, Toshiaki ; Ninomiya, Masaki ; Ito, Etsuro. / Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling. In: PLoS One. 2015 ; Vol. 10, No. 6.
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