家禽是弯曲杆菌的主要宿主,美国肠胃炎的主要原因。目前,使用选择性琼脂的选择性电镀方法,CampyCifex和改良木炭头孢哌酮脱氧胆酸盐琼脂,优先用于弯曲杆菌属的定量。在家禽产品中。由于弯曲杆菌的特殊性质,这种方法并不敏感,这可能导致检测和量化结果的偏差。因此,迫切需要弯曲杆菌检测和定量方法。目的是使用BAX®SystemReal-TimePCR检测弯曲杆菌,开发一种缩短的基于富集的定量方法,用于冷藏后家禽淋洗液中的弯曲杆菌(CampyQuant™)。空肠弯曲杆菌检测的特异性和敏感性,大肠杆菌,并测定纯培养物中的拉氏梭菌。BAX®SystemReal-TimePCR测定在4.00至9.00Log10CFU/mL的计数范围内100%地一致地检测和鉴定每个物种。阐明了使用BAX®SystemReal-TimePCR测定的低水平浓度(0.00、1.00和2.00Log10CFU/mL)的弯曲杆菌的富集时间参数。确定需要20小时的富集时间来检测至少1.00Log10CFU/mL的弯曲杆菌属。使用BAX®System实时PCR测定弯曲杆菌。因此,检测时间,检测限,和富集参数用于使用来自BAX®系统实时PCR测定的检测样品来形成CampyQuant™线性标准曲线,以定量冷藏后家禽漂洗液中的水平。使用来自BAX®System实时PCR测定的循环阈值为每个弯曲杆菌属物种生成线性拟合方程,以估计检测到的洗脱液1.00至4.00Log10CFU/mL的预富集。每个方程的统计分析得出的R2为0.93、0.76和0.94,空肠弯曲杆菌的Log10RMSE为0.64、1.09和0.81,大肠杆菌,还有C.Lari,分别。该研究表明,BAX®SystemReal-TimePCR检测弯曲杆菌的速度更快,准确,弯曲杆菌计数的有效替代方法。
Poultry is the primary reservoir of Campylobacter, a leading cause of gastroenteritis in the United States. Currently, the selective plating methodology using selective agars, Campy Cefex and Modified Charcoal Cefoperazone Deoxycholate agar, is preferentially used for the quantification of Campylobacter spp. among poultry products. Due to the specific nature of Campylobacter, this methodology is not sensitive, which can lead to skewed detection and quantification results. Therefore, Campylobacter detection and quantification methods are urgently needed. The objective was to develop a shortened enrichment-based quantification method for Campylobacter (CampyQuant™) in post-chill poultry rinsates using the BAX® System Real-Time PCR assay for Campylobacter. The specificity and sensitivity for the detection of C. jejuni, C. coli, and C. lari in pure culture were determined. The BAX® System Real-Time PCR assay consistently detected and identified each species 100% of the time with an
enumeration range of 4.00 to 9.00 Log10 CFU/mL. Enrichment time parameters for low-level concentrations (0.00, 1.00, and 2.00 Log10 CFU/mL) of Campylobacter using the BAX® System Real-Time PCR assay were elucidated. It was determined that an enrichment time of 20 h was needed to detect at least 1.00 Log10 CFU/mL of Campylobacter spp. Using the BAX® System Real-Time PCR assay for Campylobacter. As a result, time of detection, detection limits, and enrichment parameters were used to develop the CampyQuant™ linear standard curve using the detected samples from the BAX® System Real-Time PCR assay to quantify the levels in post-chill poultry rinsates. A linear fit equation was generated for each Campylobacter species using the cycle threshold from the BAX® System Real-Time PCR assay to estimate a pre-enrichment of 1.00 to 4.00 Log10 CFU/mL of rinsates detected. The statistical analyses of each equation yielded an R2 of 0.93, 0.76, and 0.94 with a Log10 RMSE of 0.64, 1.09, and 0.81 from C. jejuni, C. coli, and C. lari, respectively. The study suggests that the BAX® System Real-Time PCR assay for Campylobacter is a more rapid, accurate, and efficient alternative method for Campylobacter
enumeration.