Rapid Detection of Human Immunodeficiency Virus Types 1 and 2 by Use of an Improved Piezoelectric Biosensor

Abstract

c Disasters can create situations in which blood donations can save lives. However, in emergency situations and when resources are depleted, on-site blood donations require the rapid and accurate detection of blood-borne pathogens, including human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2). Techniques such as PCR and antibody capture by an enzyme-linked immunosorbent assay (ELISA) for HIV-1 and HIV-2 are precise but time-consuming and require sophisticated equipment that is not compatible with emergency point-of-care requirements. We describe here a prototype biosensor based on piezoelectric materials functionalized with specific antibodies against HIV-1 and HIV-2. We show the rapid and accurate detection of HIV-1 and HIV-2 in both simple and complex solutions, including human serum, and in the presence of a cross-confounding virus. We report detection limits of 12 50% tissue culture infective doses (TCID 50 s) for HIV-1 and 87 TCID 50 s for HIV-2. The accuracy, precision of measurements, and operation of the prototype biosensor compared favorably to those for nucleic acid amplification. We conclude that the biosensor has significant promise as a successful point-of-care diagnostic device for use in emergency field applications requiring rapid and reliable testing for blood-borne pathogens. It is readily appreciated that in disaster situations, the efficiency of the emergency medical response can be greatly hampered by factors that can overwhelm or eliminate medical care resources, such as having a larger-than-expected number of trauma victims in need of untainted blood products or, on the positive side, having a larger-than-expected number of donors whose blood needs to be tested rapidly for the presence of potential blood-borne pathogens, including HIV (1). However, guidelines for which technologies are most reliable in such situations are lacking, and sophisticated laboratory equipment and aseptic conditions for the detection of HIV are not amenable for use in emergency situations, such as on-site medical care sites or rudimentary field laboratories and hospitals. Thus, there is a great need for robust, simple, reliable, and rapid point-of-care detection devices for emergency conditions (1, 2).Typical detection methods for the diagnosis of blood-borne pathogens, such as HIV, as established widely in medical laboratories, include the enzyme-linked immunosorbent assay (ELISA) and nucleic acid amplification by PCR. However, ELISAs and PCRs require specific reagents, such as special buffers and enzymes, and sophisticated, large, and costly pieces of equipment, not all of which are amenable to field application. Furthermore, analyte preprocessing is often necessary for obtaining optimal results with these technologies (3-6). In contrast, mass detection devices based on piezoelectric materials capable of generating surface acoustic waves (SAW) that can be applied to biological samples have long been recognized to yield relatively simple, robust, and rapid measurements in a real-time mode (4, 6-10). In p...

Point of care diagnostics for sexually transmitted infections: perspectives and advances

“…The future for the development of new inexpensive and accurate POCs will depend on the commitment of public health officials and industry to successfully partner to bring such assays, such as those based on chip technology and biosensors, [145–149], through regulatory requirements and into actual use in the USA and in resource-limited settings. The POC test pipeline should be filled with new POC assays in the next 5 years.…”

Rapid Detection of Ebola Virus with a Reagent-Free, Point-of-Care Biosensor

“…While the predominant species in the inactivated virus sample are disrupted particles, the biosensor phase shift response is likely to be much greater for an infectious Ebola sample containing intact virus. Based on our previous study of HIV detection [ 16 ] and the relative masses of intact HIV and Ebola [ 18 , 19 ], we would estimate a 5–10× greater sensitivity for intact Ebola virus compared to intact HIV. As a first approximation, we assume a log-linear correlation between mass and phase shift.…”

“…Antibodies were individually patterned on sensing lanes as previously described [ 16 ]. Briefly, sensor chips were coated with toluene and 3-glycidyloxypropyl trimethoxysilane (90%/10%) in an oven at 60 °C for 1.5 h and then rebaked at 100 °C for 1 h. Each lane was coated with antibodies at a concentration of 10 µg/mL in phosphate-buffered saline (PBS).…”

Biosensors for waterborne viruses: Detection and removal

“…Liu et al [60] successfully developed single-domain antibodies against bacteriophage MS2 and its coat protein, by selecting from an antibody library from llamas immunised with purified coat protein, and a second library from animals immunised with bacteriophage MS2. A range of monoclonal and polyclonal antibodies have been produced for viruses detection in recent years and applied in biosensor technology [61][62][63]. Aptamers are synthetic DNA or RNA molecules specifically designed to bind a target analyte of interest.…”