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Comparative Analysis of Cultural Isolation and Pcr Based Assay for Detection of Campylobacter Jejuni In Food and Faecal Samples

机译:食品和粪便样品中空肠弯曲菌的文化分离和基于Pcr的检测方法的比较分析

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摘要

In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43), cattle diarrheic faeces (48) and poultry faecal swabs (52) only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30), milk (35), cheese (30), only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each) as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.
机译:在本研究中,测试了基于空肠弯曲杆菌mapA基因的聚合酶链反应(PCR)的功效,用于检测自然感染的以及人和动物来源的粪便和食物样品中的空肠弯曲菌。同时,对所有样品进行生物体的培养分离和生化表征。阳性样品在PCR分析中扩增了一个〜589 bp大小的DNA片段,而从大肠杆菌,李斯特菌,沙门氏菌和葡萄球菌中提取的DNA中没有此类扩增子,证实了引物的特异性。随机采集的143份粪便样本包括人腹泻粪便(43),牛腹泻粪便(48)和家禽粪便拭子(52)分别只能分别检测到4、3和8,而分别检测出6、3和10。通过PCR发现为阳性。但是,在食物样本中。牛肉(30),牛奶(35),奶酪(30),通过培养和PCR检测均仅检测到一份牛肉样品。此外,相对于可以分别检测86.7%和80%样品中微生物的培养物分离,发现加标粪便和食物样品(每个样品96.1%)中PCR对空肠弯曲杆菌的检测更为敏感。结果表明,由于其高灵敏度,特异性和自动化潜力,PCR在快速筛选样品方面具有卓越的功效。

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