Simon Bystryak1, Chitrangada Acharya2. 1. Allied Innovative Systems, 1 Jill Court, Bldg. 16, Unit 2, Hillsborough, NJ 08844, United States. Electronic address: sbystryak@allisystems.com. 2. Allied Innovative Systems, 1 Jill Court, Bldg. 16, Unit 2, Hillsborough, NJ 08844, United States.
Abstract
BACKGROUND: We describe a photochemical signal amplification method (PSAM) for increasing the sensitivity of enzyme-linked immunosorbent assays (ELISAs) for the detection of HIV-1 p24 antigen, and present a preliminary validation study on ELISA+PSAM technology for detection of HIV-1 p24 antigen in clinical samples. METHODS: ELISA+PSAM is compatible with commercially available microtiter plate readers, employs an inexpensive illumination device and the amplification takes around 10 min. RESULTS: The PSAM technology not only increases the analytical sensitivity for detection of HIV-1 p24 antigen by approximately 40 times, but also significantly increases the clinical sensitivity of the ELISA: in instances where viral RNA load is <3000 copies/ml, conventional heat mediated immune complex disruption ELISA (HM-ELISA) cannot detect any HIV positive samples whereas HM-ELISA+PSAM can detect HIV infection in approximately half of the samples (clinical sensitivity is 52.63%). For viral RNA loads between 3000 and 30,000 copies/ml, the clinical sensitivities of the HM-ELISA and HM-ELISA+PSAM are 32.6% and 91.3%, and for that >30,000 copies/ml, clinical sensitivities of HM-ELISA and HM-ELISA+PSAM are 52.3% and 100%, respectively. CONCLUSIONS: The HM-ELISA+PSAM represents an advancement in monitoring HIV-1 disease progression and treatment in the global healthcare setting.
BACKGROUND: We describe a photochemical signal amplification method (PSAM) for increasing the sensitivity of enzyme-linked immunosorbent assays (ELISAs) for the detection of HIV-1p24 antigen, and present a preliminary validation study on ELISA+PSAM technology for detection of HIV-1p24 antigen in clinical samples. METHODS: ELISA+PSAM is compatible with commercially available microtiter plate readers, employs an inexpensive illumination device and the amplification takes around 10 min. RESULTS: The PSAM technology not only increases the analytical sensitivity for detection of HIV-1p24 antigen by approximately 40 times, but also significantly increases the clinical sensitivity of the ELISA: in instances where viral RNA load is <3000 copies/ml, conventional heat mediated immune complex disruption ELISA (HM-ELISA) cannot detect any HIV positive samples whereas HM-ELISA+PSAM can detect HIV infection in approximately half of the samples (clinical sensitivity is 52.63%). For viral RNA loads between 3000 and 30,000 copies/ml, the clinical sensitivities of the HM-ELISA and HM-ELISA+PSAM are 32.6% and 91.3%, and for that >30,000 copies/ml, clinical sensitivities of HM-ELISA and HM-ELISA+PSAM are 52.3% and 100%, respectively. CONCLUSIONS: The HM-ELISA+PSAM represents an advancement in monitoring HIV-1 disease progression and treatment in the global healthcare setting.
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