TY - JOUR
T1 - Detection of HIV-1 p24 at attomole level by ultrasensitive ELISA with Thio-NAD cycling
AU - Nakatsuma, Akira
AU - Kaneda, Mugiho
AU - Kodama, Hiromi
AU - Morikawa, Mika
AU - Watabe, Satoshi
AU - Nakaishi, Kazunari
AU - Yamashita, Masakane
AU - Yoshimura, Teruki
AU - Miura, Toshiaki
AU - Ninomiya, Masaki
AU - Ito, Etsuro
N1 - Publisher Copyright:
© 2015 Nakatsuma et al.
PY - 2015/6/22
Y1 - 2015/6/22
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0131319
DO - 10.1371/journal.pone.0131319
M3 - Article
C2 - 26098695
AN - SCOPUS:84939188522
SN - 1932-6203
VL - 10
JO - PloS one
JF - PloS one
IS - 6
M1 - e0131319
ER -
