Quantification ofCampylobacterspp. in chicken carcass rinse by real-time PCR

Abstract

Aims: In this study, a real‐time quantitative polymerase chain reaction (PCR) method was examined for its ability to quantify Campylobacter spp. in chicken carcass rinses and compared with bacteriological culturing. Methods and Results: The linearity of the real‐time PCR quantification protocol was assessed on pure DNA. The amplification efficiency was 100% and the square regression coefficient (R2) was 0·998. Quantification was linear over at least 7 log units. Using a crude cell lysate gave the highest sensitivity and the detection limit of the method was 3·3 log CFU per carcass. The statistical correlation between the bacteriological enumeration and the real‐time quantitative (Q)‐PCR determined using chicken carcasses sampled at the end of the slaughter line was 0·733. The difference in detection levels was probably because of the detection of stressed, dead or viable but not culturable cells by Q‐PCR. Conclusion: The real‐time Q‐PCR method described in this study is a powerful tool for determining the number of Campylobacter cells on carcasses. Significance and Impact of the Study: The real‐time Q‐PCR method is available to quantify the Campylobacter contamination at the slaughterhouse level and could be used to evaluate primary production.

Detection of Campylobacter species in stool specimens from patients with symptoms of acute flaccid paralysis in South Africa

“…Given that gastroenteritis due to C. jejuni infections are hyperendemic among young children (< 5 years) in tropical resource-poor settings, it is expected that the prevalence of C. jejuni-induced GBS would be prevalent among individuals of < 5 years of age. Other researchers, using a broader case definition that includes all age groups for GBS surveillance, have published reports of bimodal distribution of GBS cases by age, with more cases identified in young adults (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) and the elderly (significantly higher) [4,16,29,30].…”

“…(Table 1) was used in our current study [25][26][27]. Multiplex qPCRs were performed in 50 µL reactions and included the following components: 25 µL TaqMan Gene Expression Master Mix (Applied Biosystems, Foster City, USA); 12 µL internal positive control (IPC) mix (Applied Biosystems, Foster City, USA; 7.5 µM mapA primer with 1.5 µM mapA probe; 15 µM ceuE primer with 3 µM ceuE probe; and 15 µM 16SrRNA primer with 3 µM 16SrRNA probe (Inqaba Technologies, Pretoria, South Africa, (primers); and Roche, Johannesburg, South Africa, probes); 7 µL template DNA; and 3 µL of deionized water.…”

Comparative Analysis and Limitations of Ethidium Monoazide and Propidium Monoazide Treatments for the Differentiation of Viable and Nonviable Campylobacter Cells

“…Therefore, PCR, qPCR, or reverse transcription-PCR (RT-PCR) methods for the detection and/or quantification of Campylobacter have been developed and validated during recent years (5,(23)(24)(25). In addition, EMA or PMA pretreatment of bacterial samples was implemented to circumvent the amplification of DNA from nonviable cells (9,15,16,26,27).…”

Campylobacteriosis: an emerging zoonosis, underdiagnosed and underreported by public health agencies in Brazil

“…The prevalence of Campylobacter sp in chickens and their carcasses is quite high, equivalent to 41.1% and 45.9% (BOTTELDOORN et al, 2008). A study carried out in Ireland, in 2008, showed that of 394 broiler cooled carcasses studied, 98% were contaminated with C. jejuni (EFSA, 2010).…”