恙虫病东方体黑龙江分离株的PCR/RFLP分析

Analysis on gene type of isolated strains of orientia tsutsugamushi from Heilongjiang area by way of PCR and RFLP

目的 鉴定恙虫病东方体黑龙江分离株的基因型。方法 采用PCR/RFLP技术对恙虫病东方体黑龙江分离株的sta5 6基因及其酶切图谱进行分析研究 ,并与国际参考株进行比较。结果 6株分离株的PCR特异性产物经HinfⅠ、HhaⅠ、RsaⅠ酶切后呈现 3种不同的RFLP图谱 ;MSAa930 1株、MSAa930 3株、MSAa930 4株及MSAp930 5株与Gilliam参考株具有相似的酶切图谱 ,但缺乏HinfⅠ的酶切位点 ,属于Gilliam型 ;MSAa930 6株与Karp参考株具有相同的酶切图谱 ,属于Karp型 ;MSAp930 2株与Kato参考株具有相同的酶切图谱 ,属于Kato型。 结论 黑龙江密山地区恙虫病东方体分离株存在Gilliam、Karp和Kato 3种型别。

Objective To determined the gene type of isolated strains of orientia tsutsugamushi(Ot) in Heilongjiang area.Methods PCR and RFLP were used to analyse their restriction maps of sta 56 gene of the isolated strains and made a comparison between the isolated stains of Ot and the international reference strains.Results The distinctive PCR products of the 6 isolated strains from Heilongjiang areas through restriction with Hinf Ⅰ,Hha Ⅰ,Rsa Ⅰ appeared three different kinds of RFLP restriction maps.The strains MSAa9301,MSAa9303,MSAa9304,MSAp9305 had the similar restriction map with reference strain Gilliam,but they did not have restriction spot of Hinf Ⅰ,may be classified into the same type;strains MSAa9306 had the same restriction map with reference strain karp,may be classified into the same type;strains MSAP9302 had the same restriction map with reference strain kato,may be classified into the same type.Conclusion At last three types:Gilliam,Karp and Kato were exsited in the separated specise six Ot of Heilongjiang area.