Development and Evaluation of Immunochromatographic Assay for Simple and Rapid Detection of Campylobacter jejuni and Campylobacter coli in Human Stool Specimens

Abstract

An immunochromatographic assay (Campy-ICA) using a newly generated single monoclonal antibody against a 15-kDa cell surface protein of Campylobacter jejuni was developed. When cell suspensions of 86 C. jejuni strains and 27 Campylobacter coli strains were treated with a commercially available bacterial protein extraction reagent and the resulting extracts were tested with the Campy-ICA, they all yielded positive results. The minimum detectable limits for the C. jejuni strains ranged from 1.8 ؋ 10 4 to 8.2 ؋ 10 5 CFU/ml of cell suspension, and those for the C. coli strains ranged from 1.4 ؋ 10 5 to 4.6 ؋ 10 6 CFU/ml of cell suspension. All 26 non-Campylobacter species tested yielded negative results with the Campy-ICA. To evaluate the ability of the Campy-ICA to detect C. jejuni and C. coli in human stool specimens, suspensions of 222 stool specimens from patients with acute gastroenteritis were treated with the bacterial protein extraction reagent, and the resulting extracts were tested with the Campy-ICA. The Campy-ICA results showed a sensitivity of 84.8% (28 of 33 specimens) and a specificity of 100% (189 of 189 specimens) compared to the results of isolation of C. jejuni and C. coli from the stool specimens by a bacterial culture test. The Campy-ICA was simple to perform and was able to detect Campylobacter antigen in a fecal extract within 15 min. These results suggest that Campy-ICA testing of fecal extracts may be useful as a simple and rapid adjunct to stool culture for detecting C. jejuni and C. coli in human stool specimens.

A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Detection of <i>Campylobacter</i> spp. in Stool Specimens and Comparison with Culture Method

“…Surprisingly sensitivity of culture methods was worse. It has been also reported that sensitivity and specificity of a lateral-flow-based immunoassay (Campy-ICA) was 84.8% and 100%, respectively [15]. Amri et al [1] reported that a multiplex PCR targeting cadF gene for genus Campylobacter, hipO gene for C. jejuni and asp gene for C. coli was utilized for their detection and identification of Campylobacter in stool specimens.…”

“…The different combinations of obtained data as represented in Figure suggest that the multiplex PCR is a rapid and more sensitive method to detect C. jejuni and C. coli strains but a combination with Skirrow's agar based culture method would increase the sensitivity. Recently, several molecular methods have been evaluated for the detection and identification of C. jejuni and C. coli with stool specimens [1,4,10,15,31]. An enzyme immunoassay-based antigen test (RidaScreen) showed sensitivity of 69% and specificity of 87% [31].…”

Lateral flow based immunobiosensors for detection of food contaminants

“…Campylobacter are aerobic Gram-negative helicalbacteria which differ from other food-borne pathogens in the way that do not reproduce within the food. Kawatsu et al (Kawatsu, Kumeda et al 2008) used the immunochromatographic assay method to analyze Campylobacter jejuni, based on a monoclonal antibody against cell surface protein (15 kDa). The detection limit of Campylobacter jejuni and Campylobacter coli ranged 1.8×10 4 to 8.2×10 5 CFU mL −1 and 1.4×10 5 to 4.6×10 6 CFU mL −1 , respectively (Kawatsu, Kumeda et al 2008).…”

“…Kawatsu et al (Kawatsu, Kumeda et al 2008) used the immunochromatographic assay method to analyze Campylobacter jejuni, based on a monoclonal antibody against cell surface protein (15 kDa). The detection limit of Campylobacter jejuni and Campylobacter coli ranged 1.8×10 4 to 8.2×10 5 CFU mL −1 and 1.4×10 5 to 4.6×10 6 CFU mL −1 , respectively (Kawatsu, Kumeda et al 2008). Furthermore, Klewitz et al (Klewitz, Gessler et al 2006) reported an immunochromatographic assay double sandwich format for detection of botulism neurotoxin D (BoNT/D) using a gold-anti BoNT/Dmonoclonal antibody conjugates (detector reagent) and an antiBoNT/D chicken polyclonal antibody.…”

Development of Immunochromatography-Based Methods for Detection of Leptospiral Lipopolysaccharide Antigen in Urine

“…We hypothesized that there are multiple epitopes in one molecule of LPS and that the capture antibody bound to one of the epitopes while the immobilized antibody bound to unoccupied epitopes. Usage of a single antibody for capture and immobilized antigens was also reported in ICG-based methods, which were successful in detecting Campylobacter antigens (38) and botulinum neurotoxin (39). This single antibody was adapted for our dipstick assay and ICG-based LFA, which were tested for sensitivity and specificity against Leptospira spp.…”