杏鲍菇定性及实时定量PCR检测中用作内参基因的s9ap的鉴定
Identification of s9ap used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of Pleurotus eryngii.
作者信息
Wei Yuanmiao, Liu Yao, Li Ling, Xiang Shuna, Zhang Hanyue, Shang Ying
机构信息
Institute of Food Science and Engineering, Kunming University of Science and Technology, 650500, Kunming, China.
出版信息
Mol Biol Rep. 2023 Jan;50(1):621-629. doi: 10.1007/s11033-022-07562-3. Epub 2022 Nov 12.
BACKGROUND
Pleurotus eryngii is a kind of edible fungi with good quality, and it is popular among consumers. At present, some adulterated edible fungi are available in the market. The rights and interests of consumers can be ensured by establishing a practical edible fungi detection system. Among the existing methods for detecting food adulteration, endogenous reference gene amplification is convenient and reliable. However, no ideal endogenous reference gene is available for P. eryngii.
METHODS AND RESULTS
In this study, s9ap was screened as an endogenous reference gene through sequence alignment. Qualitative and quantitative PCR analysis of this gene was carried out in one P. eryngii variety and 18 other species. The detection limit of quantitative PCR was 400 pg, and no s9ap amplification products were detected in the 18 other species.
CONCLUSIONS
This study confirmed that s9ap was an ideal endogenous reference gene for the detection of P. eryngii. This method was also suitable for processed food products.
背景
杏鲍菇是一种品质优良的食用菌，深受消费者喜爱。目前市场上存在一些掺假的食用菌。通过建立实用的食用菌检测系统可以保障消费者的权益。在现有的食品掺假检测方法中，内源性参考基因扩增方便可靠。然而，目前还没有适用于杏鲍菇的理想内源性参考基因。
方法与结果
本研究通过序列比对筛选出s9ap作为内源性参考基因。对该基因在一个杏鲍菇品种和其他18个物种中进行了定性和定量PCR分析。定量PCR的检测限为400 pg，在其他18个物种中未检测到s9ap扩增产物。
结论
本研究证实s9ap是检测杏鲍菇的理想内源性参考基因。该方法也适用于加工食品。
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