Rapid Detection of
 Campylobacter coli
 ,
 C. jejuni
 , and
 Salmonella enterica
 on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

Yang, Hong; Berrang, M. E.; Liu, Tongrui; Hofacre, Charles L.; Sánchez, Susan; Wang, Lihua; Maurer, John J.

doi:10.1128/aem.69.6.3492-3499.2003

Cited by 68 publications

(49 citation statements)

References 59 publications

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“…With conventional PCR, Campylobacter assay sensitivities of 35 to 120 CFU/ml of chicken dung (42), 36 CFU/ml of chicken fecal suspension (34), and 10 4 CFU/g of bovine feces (20) in direct PCR have been obtained. In a PCR ELISA, a sensitivity of 40 CFU/ml of chicken carcass rinse water was obtained for C. jejuni and C. coli (17). Thus, the sensitivity of the real-time PCR assay was comparable to both other real-time assays and conventional PCR assays.…”

Section: Discussionmentioning

confidence: 75%

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Lund¹,

Nordentoft²,

Pedersen³

et al. 2004

J Clin Microbiol

180

119

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A real-time PCR assay for detecting thermophilic Campylobacter spp. directly in chicken feces has been developed. DNA was isolated from fecal material by using magnetic beads followed by PCR with a prealiquoted PCR mixture, which had been stored at ؊18°C. Campylobacter could be detected in less than 4 h, with a detection limit of 100 to 150 CFU/ml, in a fecal suspension. A bacterial internal control was added before DNA extraction to control both DNA isolation and the presence of PCR inhibitors in the samples. The assay was performed on 111 swab samples from a Danish surveillance program and compared to conventional culturing using selective enrichment. There was no statistically significant difference in performance between real-time PCR and culture by selective enrichment, and the diagnostic specificity was 0.96 with an agreement of 0.92. Therefore, the assay should be useful for screening poultry flocks for the presence of Campylobacter.Thermophilic Campylobacter spp. are the major cause of bacterial enteric infection in humans in Denmark, and the incidence of infection has been increasing in all industrialized countries, particularly since 1990 (1). The reason for this dramatic rise in the number of human cases of campylobacteriosis is not known, but poultry is often implicated as a main source of human infections, due to the high prevalence of Campylobacter in broilers. Approximately 42% of all Danish broiler flocks harbored Campylobacter at the point of slaughter in 2002 (2).As food safety has become an increasing concern for consumers, there is a growing need for fast and sensitive methods for specific detection and identification of zoonotic microorganisms. The PCR technique has several advantages over classical bacteriology with respect to detection limit, speed, and the potential for automation and has succesfully been applied to the detection of Campylobacter spp. (27,28,32,37,52).However, the conventional PCR technique has certain drawbacks. In most analyses, the PCR products are separated on an agarose gel and vizualised on a UV board, with the size of the vizualised bands as the sole confirmation of specificity (12,28). Further specificity can be added by performing hybridization using blotting of PCR products (13, 51) or a PCR enzymelinked immunosorbent assay (ELISA) procedure (9, 15), but both are laborious and time-consuming processes.A disadvantage in the diagnostic application of conventional PCR is the production of false-positive results. Theese are attributable to contamination by nucleic acids, particularly from previously amplified material (carryover) (25). Any contaminant, even the smallest airborne remnant carried over from the previous PCR procedure, may be multiplied and produce a false-positive result. The real-time mode of amplification has abolished the need to open the PCR tubes following amplification and thereby has drastically reduced the risk of carryover contamination. In addition, the liquid hybridization assay (e.g., TaqMan probes) adds further specificity to the system, c...

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Section: Discussionmentioning

confidence: 75%

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Lund¹,

Nordentoft²,

Pedersen³

et al. 2004

J Clin Microbiol

180

119

View full text Add to dashboard Cite

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“…Detection limits determined in studies reporting LODs based on postenrichment cell densities are in the range of 10 3 to 10 6 CFU/ml (5,6,17), comparable to the LOD determined for this study. A PCR system able to detect Salmonella in a carcass rinse sample without pre-enrichment has recently been described (30); the LOD reported (10 2 to 10 4 CFU/cm 2 ) is comparable to that obtained for the MAAB (2.7 ϫ 10 3 CFU/ cm 2 ). The issue of cross-reactivity was addressed by performing assays in the presence of two other pathogens often found in the same types of foods.…”

Section: Discussionmentioning

confidence: 97%

Detection ofSalmonella entericaSerovar Typhimurium by Using a Rapid, Array-Based Immunosensor

Taitt

Shubin

Angel

et al. 2004

Appl Environ Microbiol

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The multianalyte array biosensor (MAAB) is a rapid analysis instrument capable of detecting multiple analytes simultaneously. Rapid (15-min), single-analyte sandwich immunoassays were developed for the detection of Salmonella enterica serovar Typhimurium, with a detection limit of 8 ؋ 10 4 CFU/ml; the limit of detection was improved 10-fold by lengthening the assay protocol to 1 h. S. enterica serovar Typhimurium was also detected in the following spiked foodstuffs, with minimal sample preparation: sausage, cantaloupe, whole liquid egg, alfalfa sprouts, and chicken carcass rinse. Cross-reactivity tests were performed with Escherichia coli and Campylobacter jejuni. To determine whether the MAAB has potential as a screening tool for the diagnosis of asymptomatic Salmonella infection of poultry, chicken excretal samples from a private, noncommercial farm and from university poultry facilities were tested. While the private farm excreta gave rise to signals significantly above the buffer blanks, none of the university samples tested positive for S. enterica serovar Typhimurium without spiking; dose-response curves of spiked excretal samples from university-raised poultry gave limits of detection of 8 ؋ 10 3 CFU/g.

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“…Rapid immunological detection and identification including latex agglutination, enzyme immunoassays (EIA) [78][79][80] ELISA [81][82][83][84] have been developed for Salmonella. There are a number of commercially available assays for Salmonella detection using anti-LPS IgM and IgG antibodies [85].…”

Section: Immune-based Rapid Detection and Identification Methodsmentioning

confidence: 99%

Recent and the Latest Developments in Rapid and Efficient Detection of Salmonella in Food and Water

Hu¹,

Li²

2017

Adv Tech Biol Med

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Salmonella is a leading cause of foodborne diseases. Gastroenteritis caused by nontyphoid Salmonella is still a major infectious disease in the world. About 95% of Salmonella infections are caused by ingestion of contaminated food and water. Rapid, sensitive, and efficient detection and identification of Salmonella from foods and water are critical for minimizing the spread of outbreaks caused by this pathogen. These methods can be applied to track the food source of contamination or by early diagnosis of the infections in a clinical setting. Culture-based methods for detection of Salmonella are laborious and time-consuming, typically taking 5-7 days to obtain a pure culture and serovar identification. In the past few decades, molecular-based technologies have greatly shortened the time for detection and identification of bacterial pathogens from food and water, and drastically increased the specificity and sensitivity of the assays. In this review, we report an update on the development of rapid and efficient methods for the detection and identification of Salmonella in food and water, focusing on the approaches for concentration of Salmonella cells and viable cell detection, as well as the advantages and drawbacks of these methods.

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Rapid Detection of Campylobacter coli , C. jejuni , and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

Cited by 68 publications

References 59 publications

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Detection ofSalmonella entericaSerovar Typhimurium by Using a Rapid, Array-Based Immunosensor

Recent and the Latest Developments in Rapid and Efficient Detection of Salmonella in Food and Water

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