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为了筛选快速、灵敏的梨火疫病菌检测方法,利用常规PCR、套式PCR和实时荧光PCR方法分别对美国进境的326批樱桃果实中梨火疫病菌进行检测。结果显示,3种PCR方法的检出率不同,不同引物或探针的检出率也存在差异。在常规PCR中,引物Ams3/Ams4c、P29A/P29B和PEANT1/PEANT2的检出率分别为35.28%、24.85%和16.87%;单管套式PCR和套式PCR的检出率分别为23.01%和50.61%;4种实时荧光PCR的检出率分别为17.48%(探针PA)、32.21%(探针Ams)、29.14%(探针ITS)和23.93%(SYBR GreenⅠ)。在所有试验方法中由引物P29A/P29B和PEANT1/PEANT2组成的套式PCR的检出率最高。检测结果证实了进境樱桃果实中存在梨火疫病菌DNA,套式PCR和常规PCR(引物Ams3/4c)可用于进境樱桃样品中梨火疫病菌的常规检测。
In order to screen the rapid and sensitive detection methods of Aspergillus bifidum, routine tests, nested PCR and real-time PCR were used to detect the susceptible strains of Aspergillus perfringens in 326 batches of cherry fruits entering the United States. The results showed that the detection rates of the three PCR methods were different, and the detection rates of different primers or probes were also different. In conventional PCR, the detection rates of primers Ams3 / Ams4c, P29A / P29B and PEANT1 / PEANT2 were 35.28%, 24.85% and 16.87%, respectively. The detection rates of single tube nested PCR and nested PCR were 23.01% and 50.61% respectively. The detection rates of the four kinds of real-time PCR were 17.48% (probe PA), 32.21% (probe Ams), 29.14% (probe ITS) and 23.93% (SYBR GreenⅠ) respectively. The highest detection rate was achieved by the nested PCR consisting of the primers P29A / P29B and PEANT1 / PEANT2 in all the test methods. The test results confirmed the presence of DNA of Aspergillus fumigatus in the incoming cherry fruit, and nested PCR and conventional PCR (primer Ams3 / 4c) were used for the routine detection of Aspergillus amyloliquefaciens in incoming cherry samples.