Whole blood based point-of-care assay for the detection of anti-pertussis toxin IgG antibodies.

Current serological diagnosis of pertussis is usually done by ELISA to determine serum specific anti-pertussis toxin (PT) IgG antibodies. However, the ELISAs are often central-laboratory based, require trained staff, and have long turnaround times. A rapid point-of-care (POC) assay for pertussis serology would aid in both diagnosis and surveillance of the disease. In this study, a quantitative lateral flow assay (LFA) with fluorescent Eu-nanoparticle reporters was used for the detection of anti-PT antibodies from whole blood. The assay was evaluated by testing overall 141 samples including 25 before and 116 one month after acellular pertussis booster vaccination. LFA results were compared to those obtained with standardized anti-PT IgG ELISAs with paired serum samples. Correlation between the assays was high (Pearson R = 0.832), and the achieved analytical sensitivity of the LFA was 29 IU/mL, which would be sufficient for clinically relevant cutoffs for determining recent infections. The paired samples, collected pre- and post-booster, demonstrated a significant increase in anti-PT IgG antibodies similar to that detected by ELISA. The developed LFA opens up several alternatives for a suitable POC test also in middle- and low-income countries.

Sensitive and quantitative detection of cardiac troponin I with upconverting nanoparticle lateral flow test with minimized interference.

Measurement of cardiac troponin I (cTnI) should be feasible for point-of-care testing (POCT) to diagnose acute myocardial infarction (AMI). Lateral flow immunoassays (LFIAs) have been long implemented in POCT and clinical settings. However, sensitivity, matrix effect and quantitation in lateral flow immunoassays (LFIAs) have been major limiting factors. The performance of LFIAs can be improved with upconverting nanoparticle (UCNP) reporters. Here we report a new methodological approach to quantify cTnI using UCNP-LFIA technology with minimized plasma interference. The performance of the developed UCNP-LFIA was evaluated using clinical plasma samples (n = 262). The developed UCNP-LFIA was compared to two reference assays, the Siemens Advia Centaur assay and an in-house well-based cTnI assay. By introducing an anti-IgM scrub line and dried EDTA in the LFIA strip, the detection of cTnI in plasma samples was fully recovered. The UCNP-LFIA was able to quantify cTnI concentrations in patient samples within the range of 30-10,000 ng/L. The LoB and LoD of the UCNP-LFIA were 8.4 ng/L and 30 ng/L. The method comparisons showed good correlation (Spearman's correlation 0.956 and 0.949, p < 0.0001). The developed UCNP-LFIA had LoD suitable for ruling in AMI in patients with elevated cTnI levels and was able to quantify cTnI concentrations in patient samples. The technology has potential to provide simple and rapid assay for POCT in ED setting.

Multiplex Point-of-Care Tests for the Determination of Antibodies after Acellular Pertussis Vaccination.

Most of the current serological diagnosis of pertussis is based on pertussis toxin (PT) IgG antibodies and does not differentiate between vaccination and infection-induced antibodies. PT is included in all of acellular pertussis vaccines available in the world. Multiplex testing of non-vaccine antigen-related antibodies has the potential to improve the diagnostic outcome of these assays. In this study, we developed a quantitatively spatial multiplex lateral flow immunoassay (LFIA) for the detection of IgG antibodies directed against PT, pertactin (PRN), and filamentous hemagglutinin (FHA). The assay was evaluated with serum samples with varying anti-PT, anti-PRN, and anti-FHA IgG levels and the result was compared to those obtained with standardized ELISA. The developed assay showed good specificity with PT and PRN antibodies and semiquantification throughout the antigen combinations. This exploratory study indicates that the multiplex LFIA is specific and sensitive, and a similar test platform with alternative antigens could be suitable for new type of pertussis serology.