家蚕微粒子病的PCR诊断技术研究

在ＤＮＡ水平上，以聚合酶链反应（ＰｏｌｙｍｅｒａｓｅＣｈａｉｎＲｅａｃｔｏｎ，ＰＣＲ）技术检测家蚕微孢子虫的结果。设计、合成了两对引物，其中引物Ⅰ是针对家蚕微泡子虫（ＮｏｓｅｍａｂｏｍｂｙｃｉｓＮ．ｂ．）引物Ⅱ是针对变形孢子虫（ＶａｉｒｉｍｏｒｐｈａｎｅｃａｔｒｉｘＶ．ｎ，）的。用这两对引物分别对“桑尺蠖微孢子虫”孢子ＤＮＡ和Ｎ．ｂ．（镇江株）的纯孢子及其感染的幼虫、蛹及蛾的ＤＮＡ进行ＰＣＲ扩增，均获得预期的阳性条带；对不同引物扩增的产物进行了ＤＮＡ序列分析。初步认为引物Ⅰ可作为家蚕微孢子虫Ｎ．ｂ．特异性较高的检测引物，而引物１是微孢子虫共有的检测引物。进一步讨论了对家蚕微孢子的检测及分类上的问题。

STUDIES ON PCR INSPECTION TECHHNIQUE OF Nosema bombycis

Two pairs of primers have been designed andsynthesized to be applied in PCR in-spection of Nosema bombycis(Zhenjiang strain).One of them was from the sequence of a putative pseudogene of small subunit rRNA(SSUrRNA)of N.b.SESNu.The other was from the sequence of a gene of SSUrRNA of Vairimorpha necatrix.DNAs from thereference strain of microsporidia.Hemerophila atrilineata,Bultler spores,and those of the larva,pupa,moth inoculated with Nosema bombycis,were specifically amplified byPCR.Both of them gave expected positive bands.We sequenced the products amplifiedwith the above primers.It was tentatively considered that primerⅠis specific to N.b.,while primer Ⅱ might be employed in all microporidia,In this report,the diagnosticmethod and classification of N.b.have also been discussed.