Forensic DNA Division, National Forensic Service, Wonju 26460, the Republic of Korea; Division of Emerging Infectious Disease Response, Bureau of Infectious Disease Emergency Preparedness and Response, Korea Disease Control and Prevention Agency, Cheongju-si, 28159, the Republic of Korea.
Forensic DNA Division, National Forensic Service, Wonju 26460, the Republic of Korea.
Forensic Sci Int. 2022 Oct;339:111416. doi: 10.1016/j.forsciint.2022.111416. Epub 2022 Aug 4.
The opium poppy acts as an important natural pain reliever but is also responsible for increased rates of severe drug abuse and addiction owing to its characteristic psychoactive effect. Non-medical illicit use of the poppy plant is markedly increasing worldwide, thereby highlighting the need for a robust species identification strategy. In this study, we identified SNPs within the region of two universal DNA barcodes, matK (maturase K) and the trnL-trnF (tRNA-Leu [3'exon]-tRNA-Phe [exon] intergenic spacer, that are forensically applicable for distinguishing opium poppy species based on a genetic analysis of 164 samples of family Papaveraceae obtained from locations spanning Jeolla-do and Jeju Island, Republic of Korea. A comparative analysis of the DNA barcode sequences for two narcotic types of the Papaver species (Papaver somniferum, Papaver somniferum subs. setigerum) to eight non-narcotic species revealed three unique nucleotide substitution events. Newly identified SNPs were located at position 255 of matK and at positions 305 and 306 of trnL-trnF; the narcotic species contained C, A, and T, whereas non-narcotic species contained T, G, and C at these positions. Phylogenetic analysis demonstrated that newly identified SNPs, which we named PsMAT255 and PsLF305/306, could be used to clearly differentiate between the narcotic and non-narcotic types of Papaver species based on the patterns of nucleotide variation. These results indicate that the nucleotide differences between the narcotic and non-narcotic species may influence genetic markers. We, therefore, developed a novel SNP-based allelic genotyping assay using the RT-PCR system that can reliably differentiate the narcotic type of the Papaver species. In summary, our findings suggest that the newly identified species-specific SNPs of both matK and trnL-trnF can be used as identification markers of narcotic Papaver species. Furthermore, a newly developed TaqMan allelic discrimination assay may be used as a practically applicable diagnostic method to survey several illicit narcotic specimens carrying the type-specific SNP.
罂粟作为一种重要的天然止痛药，但也由于其特征性的精神活性作用而导致严重药物滥用和成瘾率上升。全球范围内，罂粟植物的非医疗性非法使用明显增加，因此需要制定强有力的物种鉴定策略。在这项研究中，我们鉴定了两个通用 DNA 条码区域内的 SNPs，即 matK（成熟酶 K）和 trnL-trnF（tRNA-Leu [3' exon]-tRNA-Phe [exon] 基因间隔区，这些 SNPs 可基于对从韩国全罗道和济州岛地区采集的 164 个罂粟科样本的遗传分析，用于区分罂粟物种。对两种罂粟属（罂粟、罂粟亚种）的两种麻醉类型与八种非麻醉类型的 DNA 条码序列进行比较分析，发现了三个独特的核苷酸取代事件。新鉴定的 SNP 位于 matK 的 255 位和 trnL-trnF 的 305 和 306 位；麻醉品种含有 C、A 和 T，而非麻醉品种含有 T、G 和 C。系统发育分析表明，新鉴定的 SNP，我们将其命名为 PsMAT255 和 PsLF305/306，可以根据核苷酸变异模式清楚地区分罂粟属的麻醉和非麻醉类型。这些结果表明，麻醉和非麻醉物种之间的核苷酸差异可能会影响遗传标记。因此，我们使用 RT-PCR 系统开发了一种基于 SNP 的等位基因基因分型检测，该方法可可靠地区分罂粟属的麻醉类型。总之，我们的研究结果表明，matK 和 trnL-trnF 中新鉴定的种特异性 SNP 可作为麻醉罂粟属物种的鉴定标记。此外，新开发的 TaqMan 等位基因鉴别检测可以作为一种实用的诊断方法，用于检测携带特定 SNP 的几种非法麻醉标本。
