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目的筛选出对孢子丝菌结合效率相对较高的探针。方法以50株纯培养的孢子丝菌及标准株为样本,以毛霉、烟曲霉、念珠菌各1株为阴性对照,用聚合酶链反应-酶联免疫法(PCR-ELISA)对已报道的5个孢子丝菌特异性探针(U26852、U26866、U26866’、M85053及AF117945)进行筛选。记录各探针与合成DNA片段杂交底物显色与否及酶免疫测定指数(EI)值。结果 PCR产物测序证实5个探针均位于产物序列上;在相同的ELISA条件下,探针U26852显色最强,EI值最高。结论针对28Sr RNA的探针U26852是目前与孢子丝菌结合效率相对较高的探针。
Objective To screen out the probes which have relatively high binding efficiency to spore-forming bacteria. Methods Fifty strains of pure-cultured sporotrichosis and standard strains were used as samples. One strain of Mucor, Aspergillus fumigatus and Candida was used as a negative control. PCR-ELISA Five sporogenes specific probes (U26852, U26866, U26866 ’, M85053 and AF117945) were screened. The hybridization of each probe with the synthetic DNA fragment was recorded and the enzyme immunoassay index (EI) was recorded. Results The sequencing of PCR products confirmed that all five probes were located in the product sequence. Under the same ELISA conditions, probe U26852 had the strongest color and the highest EI value. Conclusions U26852, a probe for 28Sr RNA, is currently a probe that binds to spores more efficiently.