高分辨率熔解曲线检测常见嗜尸性丽蝇

Identification of common necrophagous Calliphoridae by high resolution melting method

目的 建立高分辨率熔解曲线(HRM)鉴定常见嗜尸性丽蝇的检测方法,以期为嗜尸性蝇类的分子鉴定提供依据和技术支持。方法 收集洛阳地区常见嗜尸性丽蝇标本26只,经形态学鉴定后,提取腿部DNA,基于COI基因进行PCR-HRM检测,对所得HRM图谱和解链温度(Tm)数据进行分析和统计,DNA测序以验证HRM检测结果。结果 根据熔解曲线形状和解链温度可将本研究中的26只嗜尸性丽蝇归属于5属8种,与形态学鉴定、DNA测序结果完全一致。结论 基于COI片段的HRM技术是一种便捷有效的检测方法,可用于嗜尸性蝇类的快速鉴定。

Objective A high resolution melting(HRM) method was established for the identification of common necrophagous Calliphoridae, which will provide a basis and technical support for the molecular identification of necrophilic flies. Methods 26 Calliphorid flies were collected and morphologically identified. Specimens’ DNA were extracted from legs for the PCR-HRM detection based on COI gene. The obtained HRM profiles and melting temperature(Tm) were analyzed, and DNA sequencing was performed to verify the results of HRM detection. Results The 26 necrophagous Calliphorid flies were classified into 8 species of 5 genera according to the shape of melting curve and melting temperature, which were consistent with results from morphological observation and DNA sequencing.Conclusions HRM detection based on COI gene is a convenient and effective method, which can be utilized for rapid identification of necrophagous flies.