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A Novel Point-of-Care BioNanoSensor for Rapid HIV Detection and Treatment Monitoring.

Rozmyslowicz T, deSa J, Lec R, Gaulton GN - J AIDS Clin Res (2015)

Bottom Line: The detection range of the BNS device for the biomarker gp120 displayed a low-end sensitivity of 6.5×10(4) HIV viral particles/ml, while using a small fluid sample (5 µl) and with a reaction time of less then 30 seconds.Performance of this device indicated that the BNS has utility for direct detection of HIV particles prior to, and independent from, antibody formation.The BNS parameters of small sample volume, compact device size, and detection sensitivity indicate that the BNS is potentially useful in the point-of-care and/or home setting for monitoring decisions regarding HIV treatment on a real-time basis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, University of Pennsylvania, School of Medicine, Philadelphia, USA.

ABSTRACT

We report here a new diagnostic approach to the direct detection of HIV in blood or other body fluids that is rapid, sensitive and potentially applicable in a point-of-care setting. The approach follows on the development of a novel BioNanoSensor (BNS) device that utilizes piezoelectric technology to detect the presence of the HIV surface glycoprotein gp120 in a nanoscale format. The detection range of the BNS device for the biomarker gp120 displayed a low-end sensitivity of 6.5×10(4) HIV viral particles/ml, while using a small fluid sample (5 µl) and with a reaction time of less then 30 seconds. Performance of this device indicated that the BNS has utility for direct detection of HIV particles prior to, and independent from, antibody formation. Accordingly, this device holds utility to monitor the status of HIV infection both early after exposure to virus as well as during chronic HIV infection. The BNS parameters of small sample volume, compact device size, and detection sensitivity indicate that the BNS is potentially useful in the point-of-care and/or home setting for monitoring decisions regarding HIV treatment on a real-time basis.

No MeSH data available.


Related in: MedlinePlus

Response of the BNS sensor to gp120 binding: (A) The sensor resonant frequency change following sequential steps of the immobilization process and sensor exposure to gp120. (B) The difference in frequency response after each successive step of the BNS reaction.
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Figure 1: Response of the BNS sensor to gp120 binding: (A) The sensor resonant frequency change following sequential steps of the immobilization process and sensor exposure to gp120. (B) The difference in frequency response after each successive step of the BNS reaction.

Mentions: An assessment of the capacity and sensitivity for the BNS device to detect HIV was first conducted by measuring the sensor wave deflection in response to binding of the major HIV surface glycoprotein gp120. In brief, the reaction was conducted using a TSM surface first coated with polyclonal sheep anti-HIV-1 and then probed with a commercial preparation of gp120. To initiate the experiment, a 5 mm diameter, 16 mm thick 100/200 MHz quartz crystal with deposited 1.5/0.7 mm diameter gold electrodes was placed in a custom fabricated sensor holder. As shown in Figure 1A, following the sensor assembly binding inflection was monitored continuously at a frequency of 100/200 MHz as a series of 5 µl blocking and sample solutions were placed in contact with the sensor. Reference measurements were first taken using TRIS buffer (starting point measurement). Antibody solution (anti-gp120 at 1 mg/ml, dissolved in PBS) was then added and incubated for 60 minutes to allow for antibody binding to the sensor surface using a standard chemi-adsorption procedure: note initial negative resonance deflection. The antibody-coated sensor was then gently rinsed with TRIS buffer (in concentration of 20 mmol/l and pH 7.6), followed by a 15-minute incubation in PBS to gain reference measurements. Bovine serum albumin (BSA) (1 mg/ml, dissolved in PBS) was then adsorbed to the sensor surface for 1 hour to block any remaining binding sites. Residual BSA was washed off in PBS, and the sensor was then probed by the addition of 5 µl of gp120 at 0.2 mg/ml (1mg total protein, dissolved in PBS), and monitored over the next 60 min. An increase in BNS frequency response of 5–8,000 Hz was consistently observed following the sequential addition of antibody, antibody/blocker, and then antibody/blocker in the presence of gp120 as shown in Figure 1B.


A Novel Point-of-Care BioNanoSensor for Rapid HIV Detection and Treatment Monitoring.

Rozmyslowicz T, deSa J, Lec R, Gaulton GN - J AIDS Clin Res (2015)

Response of the BNS sensor to gp120 binding: (A) The sensor resonant frequency change following sequential steps of the immobilization process and sensor exposure to gp120. (B) The difference in frequency response after each successive step of the BNS reaction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4596080&req=5

Figure 1: Response of the BNS sensor to gp120 binding: (A) The sensor resonant frequency change following sequential steps of the immobilization process and sensor exposure to gp120. (B) The difference in frequency response after each successive step of the BNS reaction.
Mentions: An assessment of the capacity and sensitivity for the BNS device to detect HIV was first conducted by measuring the sensor wave deflection in response to binding of the major HIV surface glycoprotein gp120. In brief, the reaction was conducted using a TSM surface first coated with polyclonal sheep anti-HIV-1 and then probed with a commercial preparation of gp120. To initiate the experiment, a 5 mm diameter, 16 mm thick 100/200 MHz quartz crystal with deposited 1.5/0.7 mm diameter gold electrodes was placed in a custom fabricated sensor holder. As shown in Figure 1A, following the sensor assembly binding inflection was monitored continuously at a frequency of 100/200 MHz as a series of 5 µl blocking and sample solutions were placed in contact with the sensor. Reference measurements were first taken using TRIS buffer (starting point measurement). Antibody solution (anti-gp120 at 1 mg/ml, dissolved in PBS) was then added and incubated for 60 minutes to allow for antibody binding to the sensor surface using a standard chemi-adsorption procedure: note initial negative resonance deflection. The antibody-coated sensor was then gently rinsed with TRIS buffer (in concentration of 20 mmol/l and pH 7.6), followed by a 15-minute incubation in PBS to gain reference measurements. Bovine serum albumin (BSA) (1 mg/ml, dissolved in PBS) was then adsorbed to the sensor surface for 1 hour to block any remaining binding sites. Residual BSA was washed off in PBS, and the sensor was then probed by the addition of 5 µl of gp120 at 0.2 mg/ml (1mg total protein, dissolved in PBS), and monitored over the next 60 min. An increase in BNS frequency response of 5–8,000 Hz was consistently observed following the sequential addition of antibody, antibody/blocker, and then antibody/blocker in the presence of gp120 as shown in Figure 1B.

Bottom Line: The detection range of the BNS device for the biomarker gp120 displayed a low-end sensitivity of 6.5×10(4) HIV viral particles/ml, while using a small fluid sample (5 µl) and with a reaction time of less then 30 seconds.Performance of this device indicated that the BNS has utility for direct detection of HIV particles prior to, and independent from, antibody formation.The BNS parameters of small sample volume, compact device size, and detection sensitivity indicate that the BNS is potentially useful in the point-of-care and/or home setting for monitoring decisions regarding HIV treatment on a real-time basis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, University of Pennsylvania, School of Medicine, Philadelphia, USA.

ABSTRACT

We report here a new diagnostic approach to the direct detection of HIV in blood or other body fluids that is rapid, sensitive and potentially applicable in a point-of-care setting. The approach follows on the development of a novel BioNanoSensor (BNS) device that utilizes piezoelectric technology to detect the presence of the HIV surface glycoprotein gp120 in a nanoscale format. The detection range of the BNS device for the biomarker gp120 displayed a low-end sensitivity of 6.5×10(4) HIV viral particles/ml, while using a small fluid sample (5 µl) and with a reaction time of less then 30 seconds. Performance of this device indicated that the BNS has utility for direct detection of HIV particles prior to, and independent from, antibody formation. Accordingly, this device holds utility to monitor the status of HIV infection both early after exposure to virus as well as during chronic HIV infection. The BNS parameters of small sample volume, compact device size, and detection sensitivity indicate that the BNS is potentially useful in the point-of-care and/or home setting for monitoring decisions regarding HIV treatment on a real-time basis.

No MeSH data available.


Related in: MedlinePlus