In the United States shrimps is the most popular seafood. Penaeidae shrimp species are known to share morphology characteristic and identification based on morphological appearance becomes more challenging when shrimp is processed into ready-to-eat products. Misrepresentation of shrimp is common in commercially available shrimp. The objective of the study was to develop a high-resolution melting (HRM) real time PCR assay targeting the 16S rRNA gene fragment for specific identification of top five penaeid species, Litopenaeus vannamei, Penaeus duorarum, Penaeus monodon, Litopenaeus setiferus and Pleoticus muelleri. The applicability of assay was evaluated using two DNA extraction kits and two real-time PCR master mixes. The HRM assay was evaluated using 43 shrimp samples and results were validated by sequencing shrimp 16S rRNA gene fragment. Assay standardized in this study formed distinct melt curve profile for each species in the normalized and differential melt curve plots. The assay using Apex qPCR 2 × GREEN master mix showed 100% sensitivity and specificity. Further, species identification results obtained by HRM assay was in complete agreement with identification achieved by 16S rRNA gene sequencing. The HRM assay developed in this study can be used as rapid, low-cost, and reliable method for the identification of abovementioned shrimp species.

在美国，虾是最受欢迎的海鲜。已知对虾科虾具有共同的形态特征，当将虾加工成即食产品时，根据形态外观进行鉴定变得更具挑战性。在市场上可买到的虾中，虾的虚假陈述很常见。该研究的目的是开发针对16S rRNA基因片段的高分辨率熔解（HRM）实时PCR检测方法，以特异性鉴定前五种对虾物种，凡纳滨对虾，对虾，对虾，斑节对虾，对虾和对虾。使用两个DNA提取试剂盒和两个实时PCR预混液评估了测定的适用性。使用43个虾样品评估HRM分析，并通过对虾16S rRNA基因片段进行测序来验证结果。在这项研究中标准化的测定法在归一化和差分熔解曲线图中为每种物种形成了独特的熔解曲线图。使用Apex qPCR 2×GREEN预混液的分析显示出100％的灵敏度和特异性。此外，通过HRM分析获得的物种鉴定结果与通过16S rRNA基因测序获得的鉴定完全吻合。在这项研究中开发的HRM分析方法可以用作鉴定上述虾类的快速，低成本和可靠的方法。