Rapid and sensitive handheld biosensor for detection of hantavirus antibodies in wild mouse blood samples under field conditions

Abstract

Hantavirus cardiopulmonary syndrome (HCPS) is an acute, life threatening viral illness that was first recognized in the spring of 1993 during an outbreak in the Four Corners region of the southwest United States. The etiologic agent, Sin Nombre (SN) virus, is a zoonotic infection that is enzootic in deer mice ( Peromyscus maniculatus). Both human and rodent infections lead to specific antibody responses, and detection of such antibodies is the mainstay of diagnosis of infection in both species. More than 300 cases of HCPS have been confirmed in 30 states and four Canadian provinces since 1993 outbreak. Forty percent have been fatal. Conventional methods for the detection of hantavirus antibodies require 4–9 h. Since HCPS can rapidly progress to death, there is a clear need for a rapid diagnostic test. Rapid diagnostic tests for the detection of hantavirus infection should be ideally be (a) quick (b) simple to carry out, (c) allowing if possible for use in field conditions. A field-deployable test would have a particular advantage for diagnosis of infection in wild rodents, such as for risk assessments around case-households, as well as potential human diagnostic applications in battlefield situations. A prototype amperometric immunosensor based on highly dispersed immunoelectrode is developed for operation in field conditions on the importance of rapid and sensitive assay for hantavirus infection in mouse blood serum. This study was initiated to simplify and develop effective procedures for screening rodent serum samples in remote areas. A sandwich scheme immunoassay has been developed and used. Naphthol formed as a result of enzymatic reduction of naphthyl phosphate in the presence of alkaline phosphatase label has been detected amperometrically. The performance of the biosensor device is evaluated by testing mice blood samples in the wild under field conditions. The results were compared with the results obtained using the strip immunoblot assay (SIA) and that obtained from Centers for Disease Control and Prevention (CDC). The overall time of analysis is as low as 25 min and statistical analysis of the data indicated promising results. The device can be easily adapted for fast analysis of other microorganisms under non-laboratory conditions, in remote locations and field operations.