Gelatin is widely used in many confectionery products such as gummy, candies, pastilles and marshmallows. However, the acceptance of gelatin in consumer goods largely depends on its animal origins. The main sources of gelatin are pig, cow and fish collagens that have differential acceptance in Halal, Kosher and Hindu foods. Currently, several simplex and duplex PCR assays are available for the determination of gelatin origins but those methods are not only time-consuming and costly but also cannot reveal all information in a single assay platform. For the first time, we documented here a tetraplex PCR assay involving eukaryotic, fish, bovine, and porcine control for the simultaneous detection of these species by a single, easy and reliable approach. The assay was optimized under pure and mixed gelatin matrices and validated by PCR product sequencing. Finally, 38 halal branded confectionery items were screened and 33, 2 and 3 of them yielded positive bands for bovine, porcine and eukaryotes, respectively. The detection limit was 0.1 to 0.001 ng DNA extracted from pure and mixed gelatins. The study is potentially useful in monitoring and certifying halal, kosher and vegetarian issues in processed foods, pharmaceuticals and cosmetics.

明胶广泛用于许多糖果产品，例如软糖，糖果，锭剂和棉花糖。但是，明胶在消费品中的接受程度很大程度上取决于其动物来源。明胶的主要来源是猪，牛和鱼的胶原蛋白，在清真食品，犹太洁食和印度教食品中具有不同的接受度。当前，有几种单工和双工PCR测定法可用于确定明胶来源，但是这些方法不仅费时，昂贵，而且无法在单个测定平台上揭示所有信息。我们首次在这里记录了涉及真核，鱼，牛和猪控制的四重PCR检测方法，可通过一种简单，可靠的方法同时检测这些物种。该方法在纯明胶和混合明胶基质下进行了优化，并通过PCR产物测序进行了验证。最后，筛选了38个清真品牌糖果，其中33个，2个和3个分别产生了牛，猪和真核生物的阳性条带。检测限为从纯明胶和混合明胶中提取的0.1至0.001 ng DNA。这项研究可能对监测和认证加工食品，药品和化妆品中的清真，犹太洁食和素食主义者问题有用。