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Enzyme-linked immunosorbent assays (ELISA) specific for anti-HSV glycoprotein G (gG) are most commonly used in the clinical diagnosis of HSV infection. But most of them are qualitative and with narrow detection ranges. A novel time-resolved fluoroimmunoassay (TRFIA) methodology was developed for the quantitative determination of HSV IgG in human serum. The assay was based on an indirect immunoassay format, and performed in 96-well microtiter plates. HSV-1 and HSV-2 were used as the coating antigens. Eu3+-labeled goat anti-(human IgG) polyclonal antibodies were used as tracers. The fluorescence intensity of each well was measured and serum HSV IgG levels quantified against a calibration curve. The detection range of the novel TRFIA was between 5 and 500 AU/mL. Assay sensitivity was 0.568 AU/mL. The intra- and inter-assay coefficients of variation were 0.59-3.63% and 3.65-6.81%, respectively. Analytical recovery, dilution tests and serum panel tests were performed using TRFIA and the results proved satisfactory. There were no statistically significant differences in sensitivity and specificity between the TRFIA and commercial ELISAs. An effective, sensitive and accurate quantitative HSV type 1 and type 2 IgG TRFIA was successfully developed and provided diagnostic value in clinical use. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

ABSTRACT We developed a luminescent terbium sensor (LTS) based on energy resonance transfer for homogeneous bioassays. The effect of temperature on photoluminescence and time-resolved fluorescence of the LTS was investigated. When the temperature was increased from 277 K to 369 K, the photoluminescence quantum yield decreased by up to 25 %, time-resolved fluorescence decreased by up to 54 %, and the lifetime shortened dramatically. Studies showed that both photoluminescence and time-resolved fluorescence quantum yields were largely recovered after samples were heated from 298 to 310, 333 or 369 K and subsequently cooled to 298 K. These results indicate that the homogeneous bioassay with LTS is sensitive to temperature and should be conducted at a constant temperature to ensure the temperature effect does not influence data and to increase the accuracy of the results. The results of this study are important for LTS applications in homogeneous bioassays. Copyright © 2012 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Enzyme-linked immunosorbent assays (ELISA) specific for anti-HSV glycoprotein G (gG) are most commonly used in the clinical diagnosis of HSV infection. But most of them are qualitative and with narrow detection ranges. A novel time-resolved fluoroimmunoassay (TRFIA) methodology was developed for the quantitative determination of HSV IgG in human serum. The assay was based on an indirect immunoassay format, and performed in 96-well microtiter plates. HSV-1 and HSV-2 were used as the coating antigens. Eu(3+)-labeled goat anti-(human IgG) polyclonal antibodies were used as tracers. The fluorescence intensity of each well was measured and serum HSV IgG levels quantified against a calibration curve. The detection range of the novel TRFIA was between 5 and 500 AU/mL. Assay sensitivity was 0.568 AU/mL. The intra- and inter-assay coefficients of variation were 0.59-3.63% and 3.65-6.81%, respectively. Analytical recovery, dilution tests and serum panel tests were performed using TRFIA and the results proved satisfactory. There were no statistically significant differences in sensitivity and specificity between the TRFIA and commercial ELISAs. An effective, sensitive and accurate quantitative HSV type 1 and type 2 IgG TRFIA was successfully developed and provided diagnostic value in clinical use., (Copyright © 2014 John Wiley & Sons, Ltd.)

We developed a luminescent terbium sensor (LTS) based on energy resonance transfer for homogeneous bioassays. The effect of temperature on photoluminescence and time-resolved fluorescence of the LTS was investigated. When the temperature was increased from 277 K to 369 K, the photoluminescence quantum yield decreased by up to 25 %, time-resolved fluorescence decreased by up to 54 %, and the lifetime shortened dramatically. Studies showed that both photoluminescence and time-resolved fluorescence quantum yields were largely recovered after samples were heated from 298 to 310, 333 or 369 K and subsequently cooled to 298 K. These results indicate that the homogeneous bioassay with LTS is sensitive to temperature and should be conducted at a constant temperature to ensure the temperature effect does not influence data and to increase the accuracy of the results. The results of this study are important for LTS applications in homogeneous bioassays., (Copyright © 2012 John Wiley & Sons, Ltd.)

Objective: To develop an amplified luminescent proximity homogeneous immunoassay (AlphaLISA) kit for the detection of human hepatitis B virus e antibody (HBeAb)., Methods: The neutralizing and competitive inhibition method was used to develop the AlphaLISA kit for detection of serum HBeAb., Results: The working range of the kit was 0.003-16 NCU/ml with a sensitivity up to 0.003 NCU/ml. The intra- and inter-assay coefficient of variation was 5.3% and 6.8%, respectively. The kit showed no cross-reaction with HBcAb, and comparison of the detection results with those of a commercially available Elecsys HBeAb kit (Roche) for 136 samples showed a correlation coefficient of 0.961., Conclusion: The AlphaLISA kit for HBeAb detection meets the clinical requirements for detection HBeAb in human serum.

Objective: To prepare a time-resolved fluoroimmunoassay (TRFIA) kit for clinical detection of IgM antibodies to hepatitis B core antigen (HBc)., Methods: Immunocapture method was used to develop the TRFIA kit for detection of the anti-HBc IgM antibodies, and the precision, cross-reactivity and sensitivity of the kit were tested with the clinical serum samples., Results: The intra- and inter-assay coefficients of variation of the TRFIA kit were 4.8%-7.2% and 7.5%-8.6%, respectively, and no cross-reactivity with anti-HBs, anti-HBc-IgG or anti-HBe was found. Comparison of the results of the TRFIA kit and enzyme-linked immunosorbent assay (ELISA) demonstrated greater sensitivity of the kit than ELISA in detecting the anti-HBc IgM antibodies in 584 serum samples. According to the detection results in 300 serum samples from healthy donors, the cutoff value of the TRFIA kit was 4.5 times of the fluorescence value of the negative control., Conclusion: This TRFIA kit for detecting anti-HBc IgM antibodies meets the demand for clinical application and can replace the ELISA kits.