Patil-Joshi Amruta, Rangaswamy B E, Apte-Deshpande Anjali
Department of Biotechnology, BIET, Davangere, Karnataka, 577004, India.
Central Dogma Pvt Ltd., A4, Gulmohar Residency, Baner Road, Baner, Pune, Maharashtra, 411045, India.
J Genet Eng Biotechnol. 2021 Mar 1;19(1):37. doi: 10.1186/s43141-020-00110-1.
The analysis of the quality of food is important to protect humans from food-borne or food-based illnesses caused by pathogens, such as bacteria, fungi, viruses, and protozoa. Rapid identification of these pathogens is critical to ensure food safety. Various detection and identification strategies exist; however, they are laborious and time consuming and hence the detection takes longer time. The aim of this study was to develop the specific and fast method for the detection of contaminants in milk.
In this study, we have developed a simple paper-based PCR method with minimum sample preparation process. The 16S rDNA universal primers were used for the detection of bacterial contaminants. LacZ primers were used for coliform detection which causes serious illness and hence their detection is crucial. ITS region primers were used for fungal detection. The most unique thing about this study is use of Whatman paper no. 1 as sample carrier material. We developed and validated the paper-based PCR method and used it for the detection of microbes and coliforms using milk as a representative sample.
We evaluated this method for its suitability in the detection of contaminant microbes using different milk samples. The paper-based method could successfully detect contaminants in the milk samples and the results were comparable to the traditional microbial detection method. The traditional microbiological method takes at least 18-20 h for detecting the presence of microbes in any sample but the developed paper-based PCR method can confirm the microbial presence in 2-3 h. This is very promising especially in the testing where sample sterility is crucial.
食品质量分析对于保护人类免受由病原体（如细菌、真菌、病毒和原生动物）引起的食源性或基于食物的疾病至关重要。快速鉴定这些病原体对于确保食品安全至关重要。存在各种检测和鉴定策略；然而，它们费力且耗时，因此检测需要更长时间。本研究的目的是开发一种用于检测牛奶中污染物的特异性快速方法。
在本研究中，我们开发了一种样品制备过程最少的简单纸质PCR方法。使用16S rDNA通用引物检测细菌污染物。使用LacZ引物检测大肠菌群，大肠菌群会导致严重疾病，因此对其检测至关重要。使用ITS区域引物检测真菌。本研究最独特之处在于使用1号沃特曼纸作为样品载体材料。我们开发并验证了纸质PCR方法，并将其用于以牛奶为代表性样品检测微生物和大肠菌群。
我们评估了该方法在使用不同牛奶样品检测污染物微生物方面的适用性。纸质方法能够成功检测牛奶样品中的污染物，结果与传统微生物检测方法相当。传统微生物学方法检测任何样品中微生物的存在至少需要18 - 20小时，但所开发的纸质PCR方法能够在2 - 3小时内确认微生物的存在。这非常有前景，特别是在样品无菌性至关重要的检测中。
