利用小鼠模型筛选孢子丝菌病快速诊断的特异性引物

To screen specific primers for the quick diagnosis of sporotrichosis using a mouse model

目的通过感染孢子丝菌的动物模型筛选文献报道的4对申克孢子丝菌特异性引物,以确定引物的敏感性和特异性。方法12只小鼠随机分成实验及对照组,实验组皮内注射申克孢子丝菌菌悬液,不同时间留取皮损组织,分别进行真菌培养和皮损组织DNA提取,PCR扩增。结果4对引物S2-R2,SSHF31-SSHR97,ITS3-SSP,SS3-SS4在感染申克孢子丝菌的小鼠皮损中均可扩增出目的条带,但引物SSHF31-SSHR97所需浓度较高,引物S2-R2的敏感性和特异性最好,与在体外培养条件下筛试的结果相同。结论针对几丁质合成酶基因I的引物S2-R2对申克孢子丝菌的敏感性和特异性最好。

Objective To screen specific and sensitive primer to detect paraffin imbedding tissue and clinical samples from animal model of Sporotrichosis. Methed Twelve mice were divided into two groups randomly. Mice of experiment group were injected with Sporotrichosis suspension intradermally. At different time remain skin slowedpulse tissues, discerning to run eumycete cultivation and extract DNA that skin slowedpulse tissues, have the PCR amplification. Result Four pairs of primers $2-R2, SSHF31-SSHR97, ITS3-SSP, SS3-SS4 all could be amplification objective straps in the rat skin of Sporotrichosis animal model, however primer SSHF31- SSHR97 need higher density, with in vitro culture condition have the same results, primer S2-R2 are the most sensitive and specific primer. Conclusion Using Sporotrichosis infected mouse animal model to study and research samples acquired the most specific, sensitive primer also to aim directly at chitin synthase gene I S2-R2, with the Sporotrihosis pure culture strain have the same consequence.