Abstract
The objective of this work was to validate the antibody-conjugated fluorescent dye-doped silica nanoparticle- (FDS-NP-) based assay for the rapid detection of Campylobacter jejuni in chicken samples using ISO16140:2003 single-laboratory validation guidelines. This assay reversed the paradigm of microbial testing. The noncultured FDS-NPs increased the fluorescent light signal of the test, not the cell numbers, and significantly reduced the detection time. Validation results showed that relative detection level (LOD) of the assay was 103 cfu/ml. The antibody-conjugated FDS-NPs were evaluated for the detection of C. jejuni in 140 chicken samples collected from slaughterhouses (50 carcass rinse water, 60 rectal swabs, and 30 viscera contents) against ISO 10272 reference method with duplex PCR confirmation. The relative accuracy (AC), relative specificity (SP), and relative sensitivity (SE) were 95.67%, 100%, and 94.87%, respectively. The inclusivity test of C. jejuni strains was 100% positive. The exclusivity test demonstrated no cross-reactivity with 32 strains of non-Campylobacter strains. The FDS-NP assay was very fast and specific, with time to result of 30 min compared with the 2–5 days required by the reference method. The noncultured FDS-NPs demonstrated comparable performance to the cultured reference method to detect C. jejuni in poultry samples. It is applicable for effective screening of poultry product category.
Highlights
Campylobacter spp. from chicken has been considered the major cause of gastrointestinal campylobacteriosis [1]
The objective of this work was to validate test kit based on fluorescent dye-doped antibody-conjugated silica nanoparticles (FDS-NPs) for the rapid detection of Campylobacter jejuni in chicken samples following the single-laboratory method validation procedure of ISO 16140:2003 standard
Reference strains of C. jejuni (ATCC 33291, ATCC 33560, and ATCC 81176) and 47 in-house isolates of Campylobacter spp. from chicken samples were used for inclusivity testing
Summary
Campylobacter spp. from chicken has been considered the major cause of gastrointestinal campylobacteriosis [1]. Outbreaks of C. jejuni were reported worldwide with chicken meats identified as implicated foods [2, 3]. In the United States, the number of patients with C. jejuni infection was as high as 1000 per 100,000 population. More than 80% of chickens are marketed without detectable Campylobacter [4]. About 65% of Campylobacter positive flocks were reported in the central and northeastern regions of Thailand [5], and 26.6% were reportedly found in the central markets. There is a chance of bacterial contamination to the finished products
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