Parihar Ranjeet, Deb Rajib, Niharika Jagna, Thakur Priyanka, Pegu Seema Rani, Sengar Gyanendra Singh, Sonowal Joyshik, Das Pranab Jyoti, Rajkhowa Swaraj, Raj Atul, Gupta Vivek Kumar
ICAR-National Research Centre on Pig, Guwahati, Assam India.
All India Institute of Hygiene and Public Health, Government of India, Kolkata, West Bengal India.
J Food Sci Technol. 2024 Aug;61(8):1516-1524. doi: 10.1007/s13197-023-05917-7. Epub 2024 Jan 9.
and are the most important food borne pathogen transmitting from animal meat and meat products. Therefore, it is vital to design an accurate and specific diagnostic tool for identifying those food-borne pathogens in animal meat and meat products. In the current study, , methicillin-resistant and sensitive (MRSA and MSSA) were simultaneously detected using a developed triplex PCR-based technique. To obtain an optimal reaction parameter, the multiplex assay was optimised by changing just one parameter while holding the others constant. Specificity of the assay was assessed using several porcine bacterial template DNA. The plasmid DNA was used to test the multiplex PCR assay's sensitivity and interference in spiked pork samples. , MRSA, and MSSA each have PCR amplified products with sizes of 335, 533, and 209 bp, respectively. The assay detects a minimum microbial load of 10 CFU/μl for all the three pathogens and can identify bacterial DNA as low as 10 ng/µl. The assay was validated employing 210 pork samples obtained from retail meat shops and slaughter houses, with MRSA, , and MSSA with the occurrence rate of 1.9%, 42.38%, and 18.1%, respectively. The rate of mixed bacterial contamination in pork meat samples examined with the developed method was 6.19%, 1.43%, 1.90%, and 1.43% for MSSA & , MRSA & , MSSA & MRSA, and , MSSA & MRSA, respectively. The developed multiplex PCR assay is quick and efficient, and it can distinguish between different bacterial pathogens in a single reaction tube.
[细菌名称1]和[细菌名称2]是来自动物肉类和肉制品的最重要的食源性病原体。因此，设计一种准确、特异的诊断工具来鉴定动物肉类和肉制品中的那些食源性病原体至关重要。在本研究中，使用一种基于三重PCR的技术同时检测了[细菌名称3]、耐甲氧西林和敏感的[细菌名称4]（MRSA和MSSA）。为了获得最佳反应参数，在保持其他参数不变的情况下仅改变一个参数来优化多重检测。使用几种猪细菌模板DNA评估该检测的特异性。使用质粒DNA测试多重PCR检测在加标猪肉样品中的灵敏度和干扰。[细菌名称3]、MRSA和MSSA的PCR扩增产物大小分别为335、533和209 bp。该检测对所有三种病原体的最低微生物载量检测限为10 CFU/μl，并且能够鉴定低至10 ng/µl的细菌DNA。采用从零售肉店和屠宰场获得的210份猪肉样品对该检测进行验证，MRSA、[细菌名称3]和MSSA的发生率分别为1.9%、42.38%和18.1%。用所开发方法检测的猪肉样品中，MSSA与[细菌名称3]、MRSA与[细菌名称3]、MSSA与MRSA以及[细菌名称3]、MSSA与MRSA的混合细菌污染率分别为6.19%、1.43%、1.90%和1.43%。所开发的多重PCR检测快速高效，并且能够在单个反应管中区分不同的细菌病原体。
