Development of an antigen-capture monoclonal antibody–based enzyme-linked immunosorbent assay and comparison with culture for detection ofSalmonella entericaserovar Enteritidis in poultry hatchery environmental samples

Abstract

An antigen-capture enzyme-linked immunosorbent assay (ELISA) was developed for use as a presumptive screening test for detection of Salmonella enterica serovar Enteritidis and other group D Salmonella in poultry hatchery environments. A mixture of 2 monoclonal antibodies that recognize different forms of the lipopolysaccharide O-antigen was used for specific detection of group D Salmonella. The performance of the ELISA was evaluated in comparison to standard Salmonella culture procedures. Culture for each sample included nonselective enrichment with buffered peptone water and primary selective enrichment and delayed secondary enrichment with both tetrathionate and Rappaport-Vassiliadis broths. One thousand fifty-seven samples were collected from poultry hatcheries over a 5-year period (received in 85 submissions), and S. Enteritidis was recovered from 106 (10%) of them. The diagnostic sensitivity and specificity of the ELISA relative to culture were 97.2% and 99.6%, respectively, on a sample basis and were both 100% on a submission basis. Delayed secondary enrichment increased the number of S. Enteritidis culture and ELISA-positive samples as compared to nonselective enrichment and primary selective enrichment by 25%. A significantly higher (P < 0.05) number of S. Enteritidis culture- and ELISA-positive results were obtained from Rappaport-Vassiliadis broth than from tetrathionate broth or buffered peptone water cultures. The results indicate that this ELISA procedure may be useful for screening poultry hatchery environmental samples for the presence of S. Enteritidis.

Magnetic Bead‐Based Immunoassay Coupled with Tyramide Signal Amplification for Detection of Salmonella in Foods

“…As the cultural methods are extremely laborious and time-consuming to confirm a positive result, it is hard to quickly respond to any potential outbreak or food contamination. Alternatively, molecular and immunological approaches have been developed and used for detection of Salmonella, including polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), immunofluorescent, microfluidic assays and biosensors (Shukla et al, 2011;Fronczek et al, 2013;Brooks et al, 2014;Park et al, 2014;Rodriguez-Lazaro et al, 2014;Wu et al, 2014;Zhai et al, 2014). These alternative methods are rapid enough for positive results to be obtained within 48-72 h, and can provide low detection limits.…”

Simultaneous detection of assembly and disassembly of multivalent HA tag and anti‐HA antibody in single in‐capillary assay

“…Up to date, a number of methods have been widely applied to investigate antigen-antibody interaction, such as surface plasmon resonance (SPR) [1][2][3], microarray [4,5], enzyme-linked immunosorbent assay (ELISA) [6][7][8], high performance size exclusion chromatography (HPSEC) [9][10][11], capillary electrophoresis (CE) [12,13] and others. As one of the most powerful micro-separation techniques, CE has advantages of high separation resolution, high sensitivity and low sample consumption.…”

A novel in‐capillary assay for dynamically monitoring fast binding between antibody and peptides using CE

“…Although several elegant techniques have been developed for the study of antigen-antibody/peptide-protein interactions, such as surface plasmon resonance [1][2][3][4][5], ELISA [6][7][8][9], isothermal calorimetry [10,11], and microarrays [12][13][14][15][16], high-performance size-exclusion chromatography [17][18][19][20] etc., they require complex and expensive instruments, hindering ease of operation and desired separation. Therefore, a simple and rapid method is urgently needed to effectively Article Related Abbreviations: FL, fluorescence detection; FAM-DYKD, FAM-DYKDHDGDYKDHDIDYKDDDDK; M2, monoclonal anti-FLAG M2 antibody detect and monitor the binding process between antibodies and peptides.…”