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CN105695457A 一种局限青霉的检测方法 有权

一种局限青霉的检测方法
摘要：
本发明提供一种局限青霉的检测方法，设计了三对LAMP引物FIP、BIP、LF、LB、F3和B3，其中FIP为5’端生物素标记引物，LF为5’端异硫氰酸荧光素FITC标记探针，配制含三对引物和样品模板的LAMP反应体系步骤，对LAMP反应体系进行扩增，然后用LFD试纸条检测，并随后进行了灵敏度和特异性评价。结果显示，应用LAMP-LFD方法对局限青霉基因组DNA检测最低检测线为4.2fg/μL；特异性验证本发明只对局限青霉(P.restrictum)呈阳性反应。
摘要（英）：

The invention provides a penicillium restrictum detection method. Three pairs of LAMP (Loop-mediated isothermal amplification) primers including FIP, BIP, LF, LB, F3 and B3 are designed, wherein the FIP refers to a 5'-end biotin labeled primer, and the LF refers to a 5'-end FITC (fluorescein isothiocyanate) labeled probe. The method includes steps of preparation of an LAMP reaction system containing the three pairs of primers and a sample template, amplification of the LAMP reaction system, detection with an LFD (lateral flow dipstick) test strip and evaluation of sensitivity and specificity. According to results, by application of the LAMP-LFD method, the lower detection limit of penicillium restrictum genomic DNA (deoxyribonucleic acid) detection is 4.2fg/microliter; by the penicillium restrictum detection method, only the penicillium restrictum is in positive reaction according to specificity verification.

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