Adulteration by Bacopa monnieri (BM) in Portulaca oleracea (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in the herbal market. The sequence divergences in internal transcribed spacer 2 (ITS2) between PO and its adulterant species were used to design diagnostic PCR primers. The specific designed primer sets were evaluated and show that the diagnostic PCR assay can be used to verify the authenticity of PO and BM. The detection limits of the primer set for PO and BM identification were 10 pg and 1 pg, respectively. The reactivity of diagnostic PCR was 0.1% PO genomic DNA and 0.01% BM genomic DNA in the test sample during DNA amplification. In addition, multiplex PCR (mPCR) for PO and BM identification was also established. The samples were more susceptible to the effect of steaming in authentication by singleplex PCR and mPCR than boiling and drying treatment. Furthermore, commercial samples from the market were used to demonstrate the applicability of the developed diagnostic PCR for PO authentication and diagnose BM adulteration, and the investigation found that approximately 72.2% (13/18) of PO plants in the herbal market were adulterated. In conclusion, the diagnostic PCR assay was successfully developed and its specificity, sensitivity and reactivity for PO and BM authentication were proven. These developed PCR-based molecular methods can be applied as an identification tool for PO authenticity and can be practically applied for inspection of BM adulteration in the herbal market in the future.

马齿苋假马齿苋(BM)掺假(PO)植物频繁出现；它会降低传统中药 (TCM) 的功效，并导致草药市场出现欺诈行为。在这项研究中，建立了一种诊断性 PCR 测定法，用于草药市场中 PO 和 BM 的快速鉴定。PO 及其掺假物种之间内部转录间隔子 2 (ITS2) 的序列差异用于设计诊断 PCR 引物。对特定设计的引物组进行了评估，表明诊断性 PCR 测定法可用于验证 PO 和 BM 的真实性。用于 PO 和 BM 鉴定的引物组的检测限分别为 10 pg 和 1 pg。诊断 PCR 的反应性是在 DNA 扩增期间测试样品中 0.1% PO 基因组 DNA 和 0.01% BM 基因组 DNA。此外，还建立了用于 PO 和 BM 鉴定的多重 PCR (mPCR)。样品在单重PCR和mPCR鉴定中比煮沸和干燥处理更容易受到蒸煮的影响。此外，来自市场的商业样品被用来证明开发的诊断 PCR 在 PO 鉴定和诊断 BM 掺假中的适用性，调查发现草药市场上大约 72.2% (13/18) 的 PO 植物被掺假。总之，成功开发了诊断性 PCR 测定法，并证明了其对 PO 和 BM 鉴定的特异性、敏感性和反应性。