J. Richard George

Future Applications of Oral Fluid Specimen Technology

Research has demonstrated that oral mucosal transudate (OMT), a serum-derived fluid that enters saliva from the gingival crevice and across oral mucosal surfaces, can be preferentially concentrated by a novel collecting system to yield detectable levels of immunoglobulins (i.e., IgG and IgM antibodies) against various bacterial and viral diseases. Assays based on OMT can aid in the diagnosis of disease and in the management of therapeutic drugs. A reliable and accurate OMT-based test to detect human immunodeficiency virus (HIV) antibodies is commercially available. Additional tests based on similar technologies may aid in the diagnosis of viral hepatitis, measles, mumps, and rubella as well as in monitoring levels of therapeutic drugs such as theophylline. The future use of OMT-based testing will likely increase because of the inherent advantages of this technology: convenience; avoidance of inadvertent transmission of blood-borne pathogens; ease of use in pediatric and geriatric populations; as well as the potential for blood-free home and workplace collection of patient samples.

Human immunodeficiency virus 1-specific Iga capture enzyme immunoassay for early diagnosis of human immunodeficiency virus 1 infection in infants

A simplified human immunodeficiency virus 1 (HIV-1)-specific IgA capture enzyme immunoassay (IgA-CEIA) was evaluated and compared with IgA-Western blot assay for early diagnosis of HIV-1 infection in infants born to seropositive women. A total of 232 coded sera collected prospectively from 70 infants were tested. All 25 sera from 10 HIV-1-negative in-

Detection of HIV Infection Using Serologic Techniques

Human immunodeficiency virus (HIV) infection, regardless of clinical stage, produces many biologic indicators of virus infection, replication, or both (Fig. 5.1). Such indicators include viremia, antibodies against viral proteins, circulating viral proteins, and nonspecific markers of infection such as neopterin, 32-microglobulins, and changes in the absolute number of and the ratio of CD4 and CD8 cells. Most of the markers can be detected and in many cases semiquantified by serologic tests (Table 5.1). These tests have been used to protect the blood supply, diagnose infections, monitor the progression of disease, monitor the efficacy of drug therapy, and diagnose infections in infants born to HIV-infected mothers. The availability of highly specific, inexpensive tests for HIV antibody, such as the enzyme immunoassay (EIA), have permitted public health agencies to conduct-large scale seroprevalence surveys to define the epidemic.

Influence of Host Factors on Immunoglobulin G Concentration in Oral Fluid Specimens

ABSTRACT The influence of host factors (tobacco use, dentition, bleeding gums, oral rinsing, nasal medications, and time since the last meal) on immunoglobulin G (IgG) concentration in oral fluids (OF) was determined by univariate and multivariate analysis. Significant differences in IgG concentration were found to be associated with human immunodeficiency virus (HIV) status (HIV antibody positive, +16.60 μg/ml, P = 0.0001), sex (female, +1.23 μg/ml, P = 0.004), dentition (+2.83 μg/ml, edentulous versus dentulous, P = 0.0001), bleeding gums (+6.35 μg/ml, P = 0.0001), and time since the last meal (+3.55 μg/ml, >6 h, P = 0.0001). These factors could impact diagnostic methods that rely on the immunoglobulin concentration in OF specimens.

Factors influencing HIV-1 banding patterns in miniaturized Western blot testing of dried blood spot specimens

In the HIV Seroprevalence Survey among Childbearing Women (SCBW), antibodies to human immunodeficiency virus type 1 are detected using enzyme immunoassays (EIA) and Western blot (WB) methods modified to accommodate samples of blood dried on special collection paper. Dried blood spot (DBS) eluates positive by EIA are tested by one of two WB methods, the miniblot technique using equipment from Immunetics Corporation and the PBS Integra assay (pageblot) from Genetic Systems. In this report we compared the performance of the two WB methods. The identity and position of the viral proteins on the WB were identified using monoclonal antibodies and monospecific antisera. The blots differed substantially in their composition and concentration of viral glycoproteins. Performance of the WB assays with DBS elution buffers from different EIA kits was equivalent except for samples eluted in the Abbott buffer, which reduced detection of antibodies to the p31, p51, p55, and p66 viral proteins. Case classification of DBS, positive sera, dilution curve samples, and seroconversion panels was equivalent by both tests in the presence of all elution buffers. Proficiency evaluation panels sent to SCBW participating laboratories over a 3-year period were used to note the differences between the two WB methods in detection of antibodies to the viral glycoproteins.

Serologic Tests for the Detection of Human Immunodeficiency Virus Infection

Persons infected with the human immunodeficiency virus (HIV) have a spectrum of clinical features, ranging from persons who are infected but appear completely healthy to those with rapid disease progression and mortality. All groups, regardless of clinical stage, possess several biologic indicators of viral infection or replication (Figure 5.1). These include viremia, antibodies against viral proteins, circulating viral proteins, and nonspecific markers such as neopterin, beta2-microglobulins, and T4 cell counts. A variety of immunologic tests currently exists for the detection of viral antibodies and protein antigens. These assays have permitted testing programs to protect the blood supply from infected units and to conduct seroprevalence surveys to define the epidemic. More recently, the presence or absence of viral and nonspecific markers have been used to predict the onset of clinical disease.

A combination immunoblot for the simultaneous detection and differentiation of HIV-1 and HIV-2

Abstract We have developed a combination immunoblot (combi-blot) for simultaneous detection and differentiation of HIV-1 and HIV-2 antibodies using HIV-1 lysate and HIV-1 and HIV-2 synthetic peptide antigens in a single assay. Minimal modification in antigen strip preparation and no modification in assay procedure of the current HIV-1 Western blot protocol was required. Implementation of this combi-blot following the use of the combination HIV-1/-2 enzyme immunoassay would simplify the current HIV testing algorithm and increase the accuracy of HIV-2 surveillance.