Search

ABSTRACT Serological assays detecting IgM antibodies in addition to IgG antibodies have a diagnostic advantage in finding early infections. Staphylococcal protein A (SpA), widely used as an antibody-detecting reagent in various immunoassays, is considered to have a high binding affinity mainly to IgG, although its interaction with other classes of immunoglobulins has also been documented. Using 28 samples from 22 HIV-1 seroconversion panels, the present study demonstrated detection of early IgM antibodies by SpA-based rapid point-of-care tests, including DPP HIV 1/2, DPP HIV-Syphilis, STAT-PAK HIV 1/2, and Sure Check HIV 1/2. Samples with predominant IgM antibodies were identified by in-house IgM assays and confirmed by pretreatment with 0.1 M 2-mercaptoethanol. Likewise, the detection of treponemal IgM antibodies was shown by DPP HIV-Syphilis assay in eight samples collected at early syphilis infection. Direct interaction between IgM and SpA immobilized in solid phase or in solution was demonstrated with purified human polyclonal IgM. A strong correlation was found between the antibody levels detected by SpA and anti-IgM reagent in the early seroconversion samples, thus supporting the evidence for IgM binding by SpA. These assays demonstrated the ability to detect IgM antibodies, which may increase test sensitivity in early infections due to a reduced serodiagnostic window. IMPORTANCE Sexually transmitted infections, including HIV and syphilis, remain a global public health concern. The main laboratory testing approach for HIV and syphilis relies on serological assays. Detection of the IgM class of antibodies may have a diagnostic advantage in finding early infections. The present study using well-characterized HIV-1 and syphilis samples has demonstrated that staphylococcal protein A employed for antibody detection in rapid point-of-care tests, including DPP HIV 1/2, DPP HIV-Syphilis, STAT-PAK HIV 1/2, and Sure Check HIV 1/2, can capture IgM antibodies in addition to IgG antibodies. The findings strongly suggest that the ability to detect IgM antibodies by these immunoassays may facilitate the identification of acute-stage HIV and syphilis infections.

ABSTRACT There is a critical need for an improved rapid diagnostic for enteric fever. We have previously demonstrated that serum IgA responses targeting Salmonella enterica serovar Typhi hemolysin E (HlyE) and lipopolysaccharide (LPS) are able to discriminate patients with acute typhoid from healthy controls in areas where enteric fever is endemic (healthy endemic controls) and from patients with other bacterial infections. We now have data demonstrating that IgA antibody responses against these antigens also work well for identifying patients with acute S. Paratyphi A infection. To develop a test for acute enteric fever detection, we have adapted a point-of-care immunochromatographic dual-path platform technology (DPP), which improves on the traditional lateral flow technology by using separate sample and conjugate paths and a compact, portable reader, resulting in diagnostics with higher sensitivity and multiplexing abilities. In this analysis, we have compared our standard enzyme-linked immunosorbent assay (ELISA) method to the DPP method in detecting acute phase plasma/serum anti-HlyE and anti-LPS IgA antibodies in a cohort of patients with culture-confirmed S. Typhi (n = 30) and Paratyphi A infection (n = 20), healthy endemic controls (n = 25), and febrile endemic controls (n = 25). We found that the DPP measurements highly correlated with ELISA results, and both antigens had an area under the curve (AUC) of 0.98 (sensitivity of 92%, specificity of 94%) with all controls and an AUC of 0.98 (sensitivity of 90%, specificity of 96%) with febrile endemic controls. Our results suggest that the point-of-care DPP Typhoid System has high diagnostic accuracy for the rapid detection of enteric fever and warrants further evaluation. IMPORTANCE Enteric fever remains a significant global problem, and control programs are significantly limited by the lack of an optimal assay for identifying individuals with acute infection. This is especially critical considering the recently released World Health Organization (WHO) position paper endorsing the role of the typhoid conjugate vaccine in communities where enteric fever is endemic. A reliable diagnostic test is needed to assess and evaluate typhoid intervention strategies and determine which high-burden areas may benefit most from a vaccine intervention. Our collaborative team has developed and evaluated a point-of-care serodiagnostic assay based on detection of anti-HlyE and LPS IgA. Our finding of the high diagnostic accuracy of the DPP Typhoid System for the rapid detection of enteric fever has the potential to have significant public health impact by allowing for improved surveillance and for control and prevention programs in areas with limited laboratory capacity.