Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus

Mycological research Pub Date : 2009-05-01 DOI:10.1016/j.mycres.2009.01.007

José Ángel Herrera-Vásquez, María del Carmen Cebrián, Ana Alfaro-Fernández, María del Carmen Córdoba-Sellés, Concepción Jordá

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引用次数: 27

Abstract

A multiplex PCR method has been developed to detect, differentiate, and confirm the morphological identification of three root infecting Olpidium spp.: O. bornovanus, O. brassicae, and O. virulentus. Of the 132 root samples examined, 101 samples were infected by Olpidium spp.. Based on the morphology of resting spores, the presence of O. bornovanus was confirmed in 20.5 % of the samples, whereas species identity could not be determined for the remaining samples because they failed to reproduce sexually. With multiplex PCR, it was possible to determine the Olpidium identity of all the infected samples, even when resting spores were not formed. This method was also effective for detecting Olpidium spp. in water samples. In addition, the specificity and sensitivity of multiplex PCR were evaluated. The multiplex PCR method was validated with samples of 9 different crops from 11 countries of America, Europe, and Africa.

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bornovanus、O. brassicae和O. virulentus同时检测和分化的多重PCR方法

采用多重PCR方法对三种感染榄属植物(O. bornovanus, O. brassicae, O. virulentus)的根进行检测、鉴别和形态鉴定。在检测的132份根样品中，有101份根样品感染了奥皮菌。根据休眠孢子的形态，20.5%的样本中证实了O. bornovanus的存在，而其余样本的物种身份无法确定，因为它们无法有性繁殖。使用多重PCR，可以确定所有感染样本的奥利匹亚菌身份，即使在休眠孢子没有形成的情况下。该方法对水样中的奥皮啶也有较好的检测效果。此外，还对多重PCR的特异性和敏感性进行了评价。用来自美洲、欧洲和非洲11个国家的9种不同作物的样品验证了多重PCR方法。

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Mycological research

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